国产三级毛片麻豆-国产精品无码一区二区三区免费-国产麻豆午夜三级精品-亚洲国产日韩?在线播放-精品国精品国产自在久国产应用-国产精品久久久中文字幕-麻豆AV无码久久精品蜜桃久久

新聞中心

News Center

當(dāng)前位置:首頁新聞中心心臟組織消化手冊

心臟組織消化手冊

更新時(shí)間:2016-02-16點(diǎn)擊次數(shù):2631

世界*實(shí)驗(yàn)材料供應(yīng)商 Worthington Biochemical 正式授權(quán)上海起發(fā)為其中國代理, Worthington Biochemical 在一直是行業(yè)的*,一直為廣大科研客戶提供zui為的產(chǎn)品和服務(wù),上海起發(fā)一直秉承為中國科研客戶帶來的產(chǎn)品,的服務(wù),簽約 Worthington Biochemical 就是為了給廣大科研客戶帶來更加完善的產(chǎn)品和服務(wù),您的滿意將是我們zui大的收獲

 Worthington Biochemical 中國代理, Worthington Biochemical 上海代理, Worthington Biochemical 北京代理,Worthington Biochemical 廣東代理, Worthington Biochemical 江蘇代理Worthington Biochemical 湖北代理,Worthington Biochemical 天津,Worthington Biochemical 黑龍江代理,Worthington Biochemical 內(nèi)蒙古代理,Worthington Biochemical 吉林代理,Worthington Biochemical 福建代理, Worthington Biochemical 江蘇代理, Worthington Biochemical 浙江代理,Worthington Biochemical 四川代理,

 

Worthington組織消化指南

引言

組織消化酶主要用于組織培養(yǎng)和細(xì)胞生物學(xué)研究的原代細(xì)胞的分離和收集。盡管這些消化酶已經(jīng)普遍應(yīng)用多年,但是對其組織消化和細(xì)胞收集的作用機(jī)制卻沒有全面的理解。  由此導(dǎo)致的結(jié)果便是選擇一種消化方法大多是隨機(jī)選擇或基于過去的經(jīng)驗(yàn),而不是在*理解消化原理的基礎(chǔ)上選擇的消化方案,以及如何根據(jù)具體實(shí)驗(yàn)?zāi)康男拚桨敢垣@得更好的實(shí)驗(yàn)結(jié)果。

細(xì)胞分離程序的目的是zui大程度的獲得功能活性的單個(gè)細(xì)胞,而影響結(jié)果的原因有很多,包括(不限于)以下:

1、組織類型

2、物種來源

3、動物年齡

4、遺傳改造情況(如knockouts

5、所用的消化培養(yǎng)液

6、所用的酶

7、消化酶中的雜質(zhì)

8、所用消化酶的工作濃度

9、溫度

10、孵育時(shí)間

通常前四個(gè)原因沒辦法選擇,而剩下的原因一般由經(jīng)驗(yàn)決定以獲取zui適實(shí)驗(yàn)結(jié)果。

通過查閱文獻(xiàn)來選擇*組織消化酶和zui適消化條件的研究人員通常被互相矛盾的實(shí)驗(yàn)數(shù)據(jù)搞得一頭霧水。這些巨大差異來源于組織胞外基質(zhì)的復(fù)雜而動態(tài)的特性,以及過去常用的粗制、未提純的組織分離酶。另外,組織胞外基質(zhì)由各種蛋白、糖蛋白、脂類和糖脂組成,這些成分的組成比例在不同物種、不同組織、甚至不同發(fā)育(年齡)之間存在明顯的差異,這些因素都極大的影響了消化方案的制定。目前通常使用的粗制酶,如鏈霉蛋白酶、NF1:250、以及含有幾種其他不同濃度蛋白酶的膠原酶、以及各種多糖酶、核酶和脂酶。

本指南總結(jié)了目前常用組織消化酶的zui適消化和細(xì)胞收集知識和方法,描述了標(biāo)準(zhǔn)的實(shí)驗(yàn)步驟, 便于制定一個(gè)*的細(xì)胞分離方案,同時(shí)列舉了一些相關(guān)的參考文獻(xiàn)。

下表列出多種組織的適宜消化酶(含參考文獻(xiàn),以組織類型和物種分類,詳見鏈接)

脂肪組織

腎上腺組織

骨組織

腦組織

軟骨組織

結(jié)腸組織

內(nèi)皮組織

上皮組織

心臟

淋巴結(jié)

乳腺

其它組織

其他組織

肌組織

神經(jīng)組織

胰腺

腮腺

垂體

前列腺

生殖

動物鱗片

皮膚

脾臟

干細(xì)胞

胸腺

甲狀腺/甲狀旁腺

扁桃體

腫瘤

 

 

 

注:使用Worthington公司消化酶的文獻(xiàn)不限于上表鏈接中所列,一般而言,Worthington公司的組織消化酶可用于大多組織類型的消化。

 

心臟組織消化手冊

 

物種詳細(xì)來源細(xì)胞名稱消化用酶培養(yǎng)基參考文獻(xiàn)
牛 HeartTrypsin: 0.06%25mM HEPES buffered Locke's solution, CMFTrifaro, J., Tang, R., and Novas, M.: Monolayer Co-Culture of Rat Heart Cells and B綿羊 Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990
(包括大鼠)Adrenal chromaffin 
 Paraneurons
CardiomyocytesCollagenase Type 2: 150 u/mlTyrode's solutionZhang,D., Wu, C., Qi, X., Meijering, R., Hoogstra-Berends, F., Tadevosyan, A., Cubukcuoglu, et al: Activation of Histone Deacetylase-6 Induces Contractile Dysfunction Through Derailment of alpha-tubulin Proteostasis in Experimental and Human Atrial Fibrillation., Circulation 129, 346-58, 2014
犬, adultAtrial and Ventricular MyocytesCollagenase Type 2: 0.065%See referenceBonilla, I., Sridhar, A., Nishijima, Y., Gyorke, S., Cardounel, A. and Carnes, C.: Differential Effects of the Peroxynitrite Donor, SIN-1, on Atrial and Ventricular Myocyte Electrophysiology., J Cardiovasc Pharmacol 61, 401-7, 2013
CardiomyocytesCollagenase Type 2: 300 u/mlTyrode's solutionXi, Y., Wu, G., Ai, T., Cheng, N., Kalisnik, J., Sun, J., Abbasi, S., Yang, D., Fan, C., Yuan, X., Wang, S. et al: Ionic Mechanisms Underlying the Effects of Vasoactive Intestinal Polypeptide on Canine Atrial Myocardium., Circ Arrhythm Electrophysiol 6, 976-83, 2013
Protease: 0.03%
Atrial MyocytesCollagenase Type 2: 0.03%Tyrode's solutionGan, T., Qiao, W., Xu, G., Zhou, X., Tang, B., Song, J., Li, Y., Zhang, J., Li, F., Mao, T. and Jiang, T.: Aging-Associated Changes in L-type Calcium Channels in the Left Atria of Dogs., Exp Ther Med 6, 919-924, 2013
CardiomyocytesCollagenase Type 2: 60 u/mlSee referenceHarleton, E., Besana, A., Comas, G., Danilo, P., Rosen, T., Argenziano, M., Rosen, M., Robinson, R. and Feinmark, S.: Ability to Induce Atrial Fibrillation in the Peri-Operative Period is Associated with Phosphorylation-Dependent Inhibition of TWIK Protein-Related Acid-Sensitive Potassium Channel 1 (TASK-1)., J Biol Chem 288, 2829-38, 2013
Protease XIV: 0.5 u/ml
Atrial and Ventricular MyocytesCollagenase Type 2: 0.05-0.08%See referenceZhang, H., Silva, J., Lin, Y., Verbsky, J., Lee, U., Kanter, E., Yamada, K., Schuessler, R. and Nichols, C.: Heterogeneity and Function of K(ATP) Channels in Canine Hearts., Heart Rhythm 10, 1576-83, 2013
犬, adultCardiomyocytesCollagenase Type 2: 0.05%HEPESCalloe, K., Nof, E., Jespersen, T., Di Diego, J., Chlus, N., Olesen, S.? Antzelevitch, C. and Cordeiro, J.: Comparison of the Effects of a Transient Outward Potassium Channel Activator on Currents Recorded from Atrial and Ventricular Cardiomyocytes., J Cardiovasc Electrophysiol 22, 1057-66, 2011
Protease Type XIV: 0.01%
犬, 20-25 kgCardiomyocytesCollagenase Type 2: 0.05%Tyrode's solutionBurashnikov, A., Zygmunt, A., Di Diego, J., Linhardt, G., Carlsson, L. and Antzelevitch, C.: AZD1305 Exerts Atrial Predominant Electrophysiological Actions and is Effective in Suppressing Atrial Fibrillation and Preventing its Reinduction in the Dog., J Cardiovasc Pharmacol 56, 80-90, 2010
犬, adultCardiomyocytesCollagenase Type 2: 0.05%M199Gavi, S., Yin, D., Shumay, E., Wang, H. and Malbon, C.: Insulin-Like Growth Factor-I Provokes Functional Antagonism and Internalization of Beta1-Adrenergic Receptors., Endocrinology 148, 2653-62, 2007
Protease: 0.008%
Atrial and Ventricular myocytesCollagenase Type 2: 0.065%Tyrode's solutionSchotten, U., de Haan, S., Verheule, S., Harks, E., Frechen, D., Bodewig, E., Greiser, M., Ram, R., Maessen, J., Kelm, M., Allessie, M. and Van Wagoner, D.: Blockade of Atrial-Specific K+-currents Increases Atrial but not Ventricular Contractility by Enhancing Reverse Mode Na+/Ca2+-Exchange.,Cardiovasc Res 73, 37-47, 2007
Atrial myocytesCollagenase Type 2: 0.013%HEPESBaba, S., Dun, W., Hirose, M. and Boyden, P.: Sodium Current Function in Adult and Aged Canine Atrial Cells., Am J Physiol Heart Circ Physiol 291, H756-61, 2006
犬, 20-30 kgCardiomyocytesCollagenase Type 2: 110 u/mlTyrode's solutionSun, H., Chartier, D., Leblanc, N. and Nat, S.: Intracellular Calcium Changes and Tachycardia-Induced Contractile Dysfunction in Canine Atrial Myocytes., Cardiovasc Res 49, 751-61, 2001
Ventricular and Atrial MyocytesCollagenase Type 4: 0.04%HEPES/Tyrode'sGintant, G.: Two Components of Delayed Rectifier Current in Canine Atrium and Ventricle. Does IKs Play a Role in the Reverse Rate Dependence of Class III Agents?, Circ Res 78, 26-37, 1996
Collagenase Type 2: 125 u/ml
Dog, mongrel, adultMyocytesHyaluronidase: 0.1%Joklik's MEMSpanier, A. and Weglicki, W.: Ca2+-Tolerant Adult Canine Myocytes: Preparation and Response to Anoxia/Acidosis, Am J Physiol 243, H448, 1982
雞, embryonicCardiomyocytesTrypsin: 0.13%HBSSBlech-Hermoni, Y. and Ladd, A.: Identification of Transcripts Regulated by CUG-BP, Elav-like Family Member 1 (CELF1) in Primary Embryonic Cardiomyocytes by RNA-seq., Genom Data 6, 74-76, 2015
Collagenase Type 2: 0.13%
Deoxyribonuclease I: 0.033%
雞 embryos, 9-11 day incubatedCardiomyocytesTrypsin: 0.25%PBSEschenhagen, T., Fink, C., Remmers, U., Scholz, H., Wattchow, J., Weil, J., et al.: Three-Dimensional Reconstitution of Embryonic Cardiomyocytes in a Collagen Matrix: A New Heart Muscle Model System, FASEB J 11, 683, 1997
小雞, embryos, 10-11 dayCardiomyocytesTrypsin: 0.17%HBSS, CMFWang, S., Greaser, M.L., Schultz, E., Bulinski, J.C., Lin, J.J., and Lessard, J.L.: Studies on Cardiac Myofibrillogenesis With Antibodies to Titin, Actin, Tropomyosin, and Myosin, J Cell Biol 107, 1075, 1988
小雞, 10 dayVentricularTrypsin: 0.025%HBSS, CMFKim, D., Okada, A., and Smith, T.W.: Control of Cytosolic Calcium Activity During Low Sodium Exposure in Cultured Chick Heart Cells, Circ Res 61, 29, 1987
小雞, embryo, 11 dayCardiomyocytesTrypsin: 0.05%HBSS, CMFJacob, R., Lieberman, M., Murphy, E., and Piwnica-Worms, D.: Effect of Sodium-Potassium Pump Inhibition and Low Sodium on Membrane Potential in Cultured Embryonic Chick Heart Cells, J Physiol 387, 549, 1987
小雞, embryonic, 11-12-day-oldMyocytesTrypsin: 0.025%CMF solutionMurphy, E., Aiton, J., Russell, C., and Lieberman, M.: Calcium Elevation in Cultured Heart Cells: Its Role in Cell Injury, Am J Physiol Cell Physiol 245 (14), C316, 1983
小雞, 8 day embryo MyocytesTrypsin NF 1:250: 0.25%Rinaldini's buffer solution, CFGross, W., Schopf-Ebner, E., and Bucher, O.: Technique for the Preparation of Homogeneous Cultures of Isolated Heart Muscle Cells, Exp Cell Res 53, 1, 1968
(also rat, neonate and mouse)
Hen, White LeghornHeartTrypsin: 0.025%Medium 199Dehann, R.: Regulation of Spontaneous Activity and Growth of Embryonic Chick Heart Cells in Tissue Culture, Dev Biol 16, 216, 1967
小雞embryos, 5 dayHeartTrypsin: 3.0%Tyrode's solution, CMFSteinberg, M.: "ECM": Its Nature, Origin, And Function in Cell Aggregation, Exp Cell Res 30, 257, 1963
Liver
貓科動物貓, mongrel, adult, 1.8-2.8 KgMyocytesCollagenase Type 2: 0.12%Kreb's Henseleit, CFSilver, L., Hemwall, E., Marino, T., and Houser, S.: Isolation and Morphology of Calcium-Tolerant Feline Ventricular Myocytes, Am J Physiol 245, H891, 1983
Ventricular
魚, 斑馬魚CardiomyocytesCollagenase Type 2: 0.5%MEMSander, V., Sune, G., Jopling, C., Morera, C. and Belmonte J.: Isolation and In Vitro Culture of Primary Cardiomyocytes from Adult Zebrafish Hearts., Nat Protoc 8, 800-9, 2013
Collagenase Type 4: 0.5%
青蛙青蛙 (Rana esculenta)MyocytesTrypsin: 0.04%CF RingerArrio-Dupont, M., and de Nay, D.: High Yield Preparation of Calcium-Tolerant Myocytes From Frog Ventricles, Biol Cell 54, 164, 1985
豚鼠Guinea pig, adultCardiomyocytesCollagenase Type 2: 100 u/mlM-199Zorn-Pauly K, Schaffer P, Pelzmann B, Bernhart E, Lang P, and Koidl B: L-type and T-type Ca2+ current in cultured ventricular guinea pig myocytes,Physiol Res 53(4), 369, 2004
豚鼠豚鼠CardiomyocytesCollagenase Type 2: 100-200 u/mlSee referenceDhamoon, A, Pandit, S., Sarmast, F., Parisian, K., Guha, P., Li, Y., Bagwe, S., Taffet, S. and Anumonwo, J.: Unique Kir2.x Properties Determine Regional and Species Differences in the Cardiac Inward Rectifier K+ Current.,Circ Res 94, 1332-9, 2004
豚鼠Guinea pig, female, 300-350gCoronary endothelialCollagenase Type 2: 0.1%see referenceBuxton I L, Kaiser R A, Oxhorn B C, Cheek D J: Evidence supporting the Nucleotide Axis Hypothesis: ATP release and metabolism by coronary endothelium, Am J Physiol Heart Circ Physiol 281, H1657-66, 2001
豚鼠Guinea pig, 250-450 gEndothelialCollagenase Type 2: 0.15%Perfusing solutionPreisig-Muller, R., Mederos Y Schnitzler, M., Derst, C., and Daut J.:Separation of Cardiomyocytes and Coronary Endothelial Cells for Cell-Specific RT-PCR, Am J Physiol 277, H413, 1999
(see reference)
豚鼠豚鼠, male, 450-600 gMyocytesHyaluronidase: 0.02%Krebs Henseleit bicarbonate bufferStemmer, P., Akera,T., Brody, T., Rardon, D., and Watanabe, A.: Isolation and Enrichment of Ca2+-Tolerant Myocytes for Biochemical Experiments from Guinea-Pig Heart, Life Sci 44, 1231, 1989
豚鼠豚鼠CardiomyocytesCollagenase: 0.04%Tyrode solution, CFIshihara, K., Mitsuiye, T., Noma, A., and Takano, M.: The Mg2+ Block and Intrinsic Gating Underlying Inward Rectification of the K+ Current in Guinea-Pig Cardiac Myocytes, J Physiol 419, 297, 1989
豚鼠豚鼠MyocytesHyaluronidase: 0.10%Bicarbonate buffer, CFBridge, J., Spitzer, K., and Ershler, P.: Relaxation of Isolated Ventricular Cardiomyocytes by a Voltage-Dependent, Science 241, 823, 1988
人類人類CardiacCollagenase: 0.045%DPBSHolt-Casper, D., Theisen, J., Moreno, A., Warren, M., Silva, F., Grainger, D., Bull, D. and Pa, A.: Novel Xeno-free Human Heart Matrix-Derived Three-Dimensional Scaffolds., J Transl Med 13, 194, 2015
Pancreatin: 0.1%
人類人類Cardiac stemCollagenase Type 2: 0.25%Joklik modified Eagle抯Avolio, E., Gianfranceschi, G., Cesselli, D., Caragnano, A., Athanasakis, E., Katare, R., Meloni, M., Palma, A., Barchiesi, A., Vascotto, C., Toffoletto, B., Mazzega, E., Finato, N., Aresu, G. and Livi, U.: Ex vivo Molecular Rejuvenation Improves the Therapeutic Activity of Senescent Human Cardiac Stem Cells in a Mouse Model of Myocardial Infarction., Stem Cells 32, 2373, 2014
人類人類Atrial myocytesCollagenase Type 1: 286 u/mlsee referencesVoigt, N., Zhou, X. and Dobrev, D.: Isolation of Human Atrial Myocytes for Simultaneous Measurements of Ca2+ Transients and Membrane Currents., J Vis Exp , e50235, 2013
Protease: 5 u/ml
人類人類MyofibersCollagenase Type 1: 0.3%see referenceAnderson, E., Rodriguez, E., Anderson, C., Thayne, K., Chitwood, W. and Kypson, A.: Increased Propensity for Cell Death in Diabetic Human Heart is Mediated by Mitochondrial-Dependent Pathways., Am J Physiol Heart Circ Physiol 300, H118, 2011
人類人類Atrial cardiomyocytesCollagenase Type 1: 286 u/mlsee referenceVoigt, N., Makary, S., Nat, S. and Dobrev, D.: Voltage-Clamp-Based Methods for the Detection of Constitutively Active Acetylcholine-Gated I(K,ACh) Channels in the Diseased Heart., Methods Enzymol 484, 653, 2010
Protease: 5 u/ml
人類人類Coronary artery smooth muscleCollagenase Type 2: 0.1%DMEMJensen, B., Swigart, P., Laden, M., DeMarco, T., Hoopes, C. and Simpson, P.: The Alpha-1D Is the Predominant Alpha-1-Adrenergic Receptor Subtype in Human Epicardial Coronary Arteries., J Am Coll Cardiol 54, 1137, 2009
Elastase: 0.05%
人類人類Smooth muscle aorticCollagenase Type 1: 0.2%DMEMMathew, S., Tustison, K., Sugatani, T., Chaudhary, L., Rifas, L. and Hruska, K.: The Mechanism of Phosphorus as a Cardiovascular Risk Factor in CKD., J Am Soc Nephrol 19, 1092, 2008
Collagenase Type 2: 0.1%
Elastase: 0.025%
人類人類 (n=16, age: 60 &plusmin;3 years)MyocardialHyaluronidase: 0.05%Medium 199Mukerjee, R., Multani, M., Sample, J., Dowdy, K., Zellner, J., Hoover, D., and Spinale, F.: Effects of adrenomedullin on human myocyte contractile function and beta-adrenergic response, J Cardiovasc Pharmacol 7 (4), 235, 2002
人類人類CardiomyocytesCollagenase Type 2: 0.05%HEPES solutionTodor Anastassia, Sharov Victor G, Tanhehco Elaine J, Silverman Norman, Bernabei Alvise, Sabbah Hani N: Hypoxia-induced cleavage of caspase-3 and DFF45/ICAD in human failed cardiomyocytes, Am J Physiol Heart Circ Physiol 283, H990-5, 2002
Collagenase Type 1: 0.025%
Protease XIV: 0.013%
人類人類HeartCollagenase Type 2: 200 u/mlTyrode's solutionHoppe, U., Jansen, E., Sudkamp, M., and Beukelmann, D.:Hyperpolarization-Activated Inward Current in Ventricular Myocytes From Normal and Failing Human Hearts, Circulation 97, 55, 1998
Myocytes
人類人類Atrial myocytesCollagenase Type 2: 0.1%see referenceVan Wagoner, D., Pond, A., McCarthy, P., Trimmer, J. and Nerbonne, J.:Outward K+ Current Densities and Kv1.5 Expression are Reduced in Chronic Human Atrial Fibrillation., Circ Res 80, 772-81, 1997
Protease Type XIV: 0.04%
人類人類, fetal, 14.5 week gestationMyocytesHyaluronidase: 0.1%HBSS with CalciumGoldman, B., and Wurzel, J.: Effects of Subc*tion and Culture Medium on Differentiation of Human Fetal Cardiac Myocytes, In Vitro Cell Dev Biol 28, 109, 1992
人類人類, both sexes, 18-81 yearsMyocardialCollagenase: 0.14%HBSS, CMFSmith, D., Glover, J., Townsend, L., and Maupin, E.: A Method for the Harvest, Culture, and Characterization of Human Adult Atrial Myocardial Cells: Correlation with Age of Donor, In Vitro Cell Dev Biol 27, 914, 1991
Atrial
人類人類, adultMyocytesTrypsin: 0.25%Joklik's MEMBugaisky, L.B.: Biology of Isolated Adult Cardiac Myocytes,Isolation and Culture of Human Adult Cardiac Myocytes,(Clark,Decker,Borg,eds), Elsevier Science Publishing Co., Inc., , 1988
人類人類Thoracic aortaCollagenase: 0.2%Phosphate buffer w/NaClHassler, O., Wiren, M., and Herbertsson, S.: The Elastic Coat of the Arterial Wall Studied with the Aid of Collagenase, Acta Pathol Microbiol Scand 57, 15, 1963
無脊椎動物E. cirhosa章魚, 260-352 克Systemic heart cardiomyocytesCollagenase: 0.025%see referenceAltimiras J., Hove-Madsen L., and Gesser H.: Ca(2+) Uptake in the Sarcoplasmic Reticulum from the Systemic Heart of Octopod Cephalopods, J Exp Biol 202, 2531, 1999
Trypsin: 0.02%
小鼠小鼠, 15-30 gmVentricular myocytesCollagenase Type 2: 0.12%see referenceHenn, M., Janjua, M., Kanter, E., Makepeace, C., Schuessler, R., Nichols, C. and Lawton, J.: Adenosine Triphosphate-Sensitive Potassium Channel Kir Subunits Implicated in Cardioprotection by Diazoxide., J Am Heart Assoc 4, , 2015
小鼠小鼠, 3-4 weekCardiac progenitorCollagenase Type 1: 0.1%DMEMWang, H., Chen, H., Feng, B., Wang, X., He, X., Hu, R., Yin, M., Wang, W., Fu, W. and Xu, Z.: Isolation and Characterization of a Sca-1+/CD31- Progenitor Cell Lineage Derived From Mouse Heart Tissue., BMC Biotech4, 75, 2014
小鼠小鼠, adultCardiac ProgenitorCollagenase Type 2: 600 u/mlMEMSantos Nascimento, D, Mosqueira, D, Sousa, L, Teixeira, M, Filipe, M, Resende, T, Araujo, A, Valente, M, Almeida, J, Martins, J, Santos, J, Barcia, R, Cruz, P, Cruz, H and Pinto-d-O, P: Human Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells Attenuate Remodeling after Myocardial Infarction by Proangiogenic, Antiapoptotic, and Endogenous Cell-Activation Mechanisms., Stem Cell Res Ther 5, 5, 2014
Deoxyribonuclease I: 60 u/ml
小鼠小鼠Cardiac progenitorCollagenase Type 2: 0.1%HBSSZafiriou, M., Noack, C., Unsold, B., Didie, M., Pavlova, E., Fischer, H., Reichardt, H., Bergmann, M., El-Armouche, A., Zimmermann, W., Zelarayan, L.: Erythropoietin Responsive Cardiomyogenic Cells Contribute to Heart Repair Post Myocardial Infarction., Stem Cells 32, 2480, 2014
Neutral Protease: 2.4 u/ml
小鼠小鼠, adultCardiomyocytesCollagenase Type 2: 0.08%see referenceKohncke, C. Lisewski, U., Schleussner, L., Gaertner, C., Reichert, S. and Roepke, T.: Isolation and Kv Channel Recordings in Murine Atrial and Ventricular Cardiomyocytes., J Vis Exp 73, e50145, 2013
小鼠小鼠CardiomyocytesCollagenase Type 2: 150 u/mlMEMHennessey, J., Wei, E. and Pitt, G.: Fibroblast Growth Factor Homologous Factors Modulate Cardiac Calcium Channels., Circ Res 113, 381-8, 2013
小鼠小鼠CardiomyocytesCollagenase Type 2: 360 u/mlL-15Touchberry, C., Green, T., Tchikrizov, V., Mannix, J., Mao, T., Carney, B., Girgis, M., Vincent, R., Wetmore, L., Dawn, B., Bonewald, L., Stubbs, J. and Wacker, M.: FGF23 is a Novel Regulator of Intracellular Calcium and Cardiac Contractility in Addition to Cardiac Hypertrophy., Am J Physiol Endocrinol Metab 304, E863, 2013
Papain: 0.4 u/ml
Deoxyribonuclease I: 40 u/ml
小鼠小鼠, 2-6 monthSinoatrial nodeCollagenase Type 2: 229 u/mlTyrode's solutionChris, C., Cardona, N., Mesirca, P., Herrmann, S., Hofmann, F., Striessnig, J., Ludwig, A., Mangoni, M. and Lee, A.: Distinct Localization and Modulation of Cav1.2 and Cav1.3 L-type Ca2+ Channels in Mouse Sinoatrial Node., J Physiol 590, 6327, 2012
Elastase: 1.9 u/ml
Protease Type XIV: 0.9 u/ml
小鼠小鼠, male, 8-25 weekCardiomyocytesCollagenase Type 2: 59 u/mlMEMCarley, A. and Kleinfeld, A.: Fatty Acid (FFA) Transport in Cardiomyocytes Revealed by Imaging Unbound FFA is Mediated by an FFA Pump Modulated by the CD36 Protein., J Biol Chem 286, 4589-97, 2011
小鼠小鼠Aortic adventitial leukocytesCollagenase Type 2: 300 u/mlPBSButcher, M., Herre, M., Ley, K. and Galkina, E.: Flow Cytometry Analysis of Immune Cells Within Murine Aortas., J Vis Exp 53, e2828, 2011
Elastase: 5.6 u/ml
小鼠小鼠, adultCardiomyocytesCollagenase Type 1: 0.1-0.4%see referenceFlynn, J., Santana, L. and Melov, S.: Single Cell Transcriptional Profiling of Adult Mouse Cardiomyocytes., J Vis Exp 58, e3302, 2011
Protease Type XIV: 0.004-.02%
小鼠小鼠, male, 6 weekCoronary endothelial cellsCollagenase Type 2: 0.1%M199Makino, A., Suarez, J., Wang, H., Belke, D., Scott, B. and Dillmann, W.:Thyroid Hormone Receptor-beta is Associated With Coronary Angiogenesis During Pathological Cardiac Hypertrophy., Endocrinology 150, 2008, 2009
Neutral Protease: 0.6 u/ml
小鼠小鼠, adultVentricular myoctesCollagenase: 0.1%M199Zhang, Y, Kanter, E., Laing, J., Aprhys, C., Johns, D., Kardami, E. and Yamada, K.: Connexin43 Expression Levels Influence Intercellular Coupling and Cell Proliferation of Native Murine Cardiac Fibroblasts., Cell Commun Adhes 15, 289, 2008
小鼠小鼠, adult, 9-13 weekCardiomyocytesCollagenase Type 2: 620 u/mlMyocyte buffer see referenceKabaeva, Z., Zhao, M. and Michele, D.: Blebbistatin Extends Culture Life of Adult Mouse Cardiac Myocytes and Allows Efficient and Stable Transgene Expression., Am J Physiol Heart Circ Physiol 294, H1667-74, 2008
Protease XIV: 0.104 u/ml
Deoxyribonuclease I: 0.0015%
小鼠小鼠, adultCardiomyocytesCollagenase: see referenceHBSSO'Connell Timothy D, Rodrigo Manoj C, Simpson Paul C: Isolation and culture of adult mouse cardiac myocytes, Methods Mol Biol 357, 271-96, 2007
小鼠小鼠Smooth muscle cellsCollagenase Type 2: 300 u/mlF10 HamVerheye, S., Martinet, W., Kockx, M., Knaapen, M., Salu, K., Timmermans, J., Ellis, J., Kilpatrick, D., DeMayer, G.: Selective Clearance of Macrophages in Atherosclerotic Plaques by Autophagy, J Am Coll Cardiol49, 706, 2007
Elastase: 5 u/ml
小鼠小鼠, adult, 6-12 weekCardiomyocytesCollagenase Type 2: 0.1%see referenceShioya, T: A Simple Technique for Isolating Healthy Heart Cells from Mouse Models., J Physiol Sci 57, 327-35, 2007
Trypsin: 0.006%
Protease XIV: 0.006%
小鼠小鼠Ventricular myocytes and mesenteric arterial SMCCollagenase Type 2: 0.06%Krebs-RingerLu Tong, Ye Dan, Wang Xiaoli, Seubert John M, Graves Joan P, Bradbury J Alyce, Zeldin Darryl C, Lee Hon-Chi: Cardiac and vascular KATP channels in rats are activated by endogenous epoxyeicosatrienoic acids through different mechanisms, J Physiol 575, 627-44, 2006
Papain: 0.175%
小鼠小鼠CardiomyocytesCollagenase Type 1: 0.17%PBSChen, H., Yong, W., Ren, S., Shen, W., He, Y., Cox, K., Zhu, W., Li, W., Soonpaa, M., Payne, RM, Franco, D., Field, L., Rosen, V., Wang, Y. and Shou, W.: Overexpression of Bone Morphogenetic Protein 10 in Myocardium Disrupts Cardiac Postnatal Hypertrophic Growth., J Biol Chem 281, 27481, 2006
小鼠小鼠, neonatalCardiomyoctesCollagenase Type 1: 150 u/mlM199Takahashi N, Wang X, Tanabe S, Uramoto H, Jishage K, Uchida S, Sasaki S, Okada Y.: ClC-3-independent sensitivity of apoptosis to Cl- channel blockers in mouse cardiomyocytes, Cell Physiol Biochem 15, 263, 2005
Trypsin: 0.01%
小鼠小鼠, 1 dayCardiomyoctesNCIS kit: per instructionsEBSSPotts, Malia B., Vaughn, Allyson E., McDonough, Holly, Patterson, Cam, and Deshmukh, Mohanish: Reduced Apaf-1 levels in cardiomyocytes engage strict regulation of apoptosis by endogenous XIAP, J Cell Biol 171, 925, 2005
小鼠小鼠Aortic endothelialCollagenase Type 2: 0.2%DMEMKobayashi, M., Inoue, K., Warabi, E., Minami, T. and Kodama, T.: A Simple Method of Isolating Mouse Aortic Endothelial Cells., J Atheroscler Thromb 12, 138, 2005
小鼠小鼠Smooth muscle cellsCollagenase Type 1: 0.1%DMEMFukumoto Yoshihiro, Deguchi Jun-o, Libby Peter, Rabkin-Aikawa Elena, Sakata Yasuhiko, Chin Michael T, Hill Christopher C, Lawler Patrick R, Varo Nerea, Schoen Frederick J, Krane Stephen M, Aikawa Masanori:Genetically determined resistance to collagenase action augments interstitial collagen accumulation in atherosclerotic plaques, Circulation 110, 1953-9, 2004
Elastase: 0.0125%
小鼠小鼠, female, 14-18 gCardiomyocytesCollagenase Type 2: 0.02%see referenceZhang Sui, Wang Dachun, Estrov Zeev, Raj Sean, Willerson James T, Yeh Edward T H: Both cell fusion and transdifferentiation account for the transformation of human peripheral blood CD34-positive cells into cardiomyocytes in vivo, Circulation 110, 3803-7, 2004
Elastase: 0.03%
Pancreatin: 0.06%
小鼠小鼠, 6-8 weekVascular smooth musclePapain: 10 u/mlEBSSQian, Q., Hunter, L., Li, M., Marin-Padilla, M., Prakash, Y., Somlo, S., Harris, P., Torres, V., and Sieck, G.: Pkd2 Haploinsufficiency Alters Intracellular Calcium Regulation in Vascular Smooth Muscle Cells, Hum Mol Genet 12(15), 1875, 2003
Elastase: .005%
Collagenase: 0.05%
Deoxyribonuclease I: 1000 u/ml
PDS kit: with modifications
小鼠小鼠CardiomyocytesCollagenase Type 2: 0.2%DMEM/F-12Rybkin, I., Markham, D., Yan, Z., Bassel-Duby, R., Williams, R., and Olson, E.: Conditional Expression of SV40 T-antigen in Mouse Cardiomyocytes Facilitates an Inducible Switch from Proliferation to Differentiation, J Biol Chem 278(18), 15927, 2003
Pancreatin: 0.06%
小鼠小鼠, neonatal 2-4 dayAtrial myocytesNCIS kit: with modifications see referenceL-15Bettahi, I,, Marker, C., Roman, M., and Wickman, K.: Contribution of the Kir3.1 Subunit to the Muscarinic-gated Atrial Potassium Channel IKACh, J Biol Chem 277, 48282, 2002
小鼠小鼠, neonatal and adultCardiomyocytesCollagenase Type 2: 0.03-0.2%Joklik's MEMSchreiber, K., Paquet, L., Allen, B., and Rindt, H.: Protein Kinase C Isoform Expression and Activity in the Mouse Heart, Am J Physiol Heart Circ Physiol281(5), H2062, 2001
小鼠小鼠, NTg and Tg, 8 - 12 weekMyocytesCollagenase Type 2: 150 u/mlJoklik's MEMNelson, D., Setser, E., Hall, D., Schwartz, S., Hewitt, T., Klevitsky, R., Osinska, H., Bellgrau, D., Duke, R., and Robbins, J.: Proinflammatory Consequences of Transgenic Fas Ligand Expression in the Heart, J Clin Invest 105 (9), 1199, 2000
小鼠小鼠, 1-6 dayCardiomyocytesCollagenase Type 2: 0.1%PBSWatzka SB, Lucien J, Shimada M, Edwards V, Yeger H, Hannigan G, and Coles JG.: Selection of viable cardiomyocytes for cell transplantation using three-dimensional tissue culture, Transplantation 70, 1310, 2000
Deoxyribonuclease I: 0.002%
小鼠小鼠, 2-4 moVentricular myocytesCollagenase Type 2: 0.05%MEMZhou YY, Wang SQ, Zhu WZ, Chruscinski A, Kobilka BK, Ziman B, Wang S, Lakatta EG, Cheng H, Xiao RP: Culture and adenoviral infection of adult mouse cardiac myocytes: methods for cellular genetic physiology, Am J Physiol Heart Circ Physiol 279, H429-36, 2000
Collagenase Type 4: 0.05%
Protease XIV: 0.002%
小鼠小鼠CardiomyocytesCollagenase Type 2: 150 u/mlJoklik's MEMChristensen G, Minamisawa S, Gruber PJ, Wang Y, Chien KR: High-efficiency, long-term cardiac expression of foreign genes in living mouse embryos and neonates, Circulation 101, 178-84, 2000
小鼠小鼠, 1-3 dayCardiomyocytesTrypsin: 0.25%FBS-MEMWang, G., and Kang, Y.: Inhibition of Doxorubicin Toxicity in Cultured Neonatal Mouse Cardiomyocytes with Elevated Metallothionein Levels, J Pharmacol Exp Ther 288(3), 938, 1999
小鼠小鼠, 1 dayCardiomyocytesNCIS kit: with modifications see referenceL-15Lader, A., Kwiatkowski, D., and Cantiello, H.: Role of Gelsolin in the Actin Filament Regulation of Cardiac L-type Calcium Channels, Am J Physiol 277, C1277, 1999
小鼠小鼠, 3-7 monthFibroblasts, mesangial, smooth muscleTrypsin: 0.25%DMEMBradshaw AD, Francki A, Motamed K, Howe C, Sage EH: Primary mesenchymal cells isolated from SPARC-null mice exhibit altered morphology and rates of proliferation, Mol Biol Cell 10, 1569-79, 1999
Collagenase: see reference
Soybean Trypsin Inhibitor: .05%
小鼠小鼠 (BALB/c), adultMyocytesCollagenase Type 2: 0.04%Tyrode's solutionFelzen, B., Shilkrut, M., Less, H., Sarapov, I., Maor, G., Coleman, R., Robinson, R., et al.: Fas (CD95/Apo-1)-Mediated Damage to Ventricular Myocytes Induced by Cytotoxic T Lymphocytes from Perforin-Deficient Mice,Circ Res 82, 438, 1998
小鼠小鼠, 1 dayCardiomyocytesNCIS kit: with modifications see referenceL-15Lader, A., Xiao, Y., Ishikawa, Y., Cui, Y., Vatner, D., Vatner, S., Homcy, C., Cantiello, H.: Cardiac Gsalpha Overexpression Enhances L-type Calcium Channels Through an Adenylyl Cyclase Independent Pathway, Proc Natl Acad Sci U S A 95, 9669, 1998
小鼠小鼠, P0-P1CardiomyocytesCollagenase: 0.045%DMEM, Medium 199Arber, S., Hunter, J., Ross, J., Hongo, M., Sansig, G., Borg, J., Perriard, J., Chien, K., and Caroni, P.: MLP-Deficient Mice Exhibit a Disruption of Cardiac Cytoarchitectural Organization, Dilated Cadiomyopathy, and Heart Failure,Cell 88, 393, 1997
小鼠小鼠, 6-9 or 21-41 dayVentricular and atrial myocytesCollagenase Type 2: 95 u/mlDMEM/TyrodesValenzuela, D., Han, X., Mende, U., Fankhauser, C., Mashimo, H., Huang, P., Pfeffer, J., Neer, E., and Fishman, M.: G alpha(o) is Necessary for Muscarinic Regulation of Ca2+ Channels in Mouse Heart, Proc Natl Acad Sci U S A 94(5), 1727, 1997
Hyaluronidase: 172.5 u/ml
Trypsin: 0.002%
Deoxyribonuclease I: 60u/ml
小鼠小鼠CardiomyocyteCollagenase: 0.17 %PBSSoonpaa, M., Kim, K., Pajak, aL., Franklin, M., and Field, L.:Cardiomyocyte DNA Synthesis and Binucleation During Murine Development,Am J Physiol 271, H2183, 1996
小鼠小鼠 embryosMyocytesCollagenase Type 1: 0.1%DMEMWobus, A., Kleppisch, T., Maltsev, V., and Hescheler, J.: Cardiomyocyte-Like Cells Differentiated In Vitro From Embryonic Carcinoma Cells P19 are Characterized by Functional Expression of Adrenoceptors and Ca2+ Channels,In Vitro Cell Dev Biol 30A, 425, 1994
小鼠小鼠 MyocytesTrypsin NF 1:250: 0.25%Rinaldini's buffer solution, CFGross, W., Schopf-Ebner, E., and Bucher, O.: Technique for the Preparation of Homogeneous Cultures of Isolated Heart Muscle Cells, Exp Cell Res 53, 1, 1968
(also rat, neonate and chick, 8 day embryo)
綿羊綿羊Atrial myocytesCollagenase Type 2: 0.055%Tyrode's solutionVoigt, N., Pearman, C., Dobrev, D. and Dibb, K.: Methods for Isolating Atrial Cells from Large Mammals and Humans., J Mol Cell Cardiol 86, 187-98, 2015
Protease XIV: 0.006%
綿羊綿羊CardiomyocytesCollagenase Type 2: 0.055%See referenceDibb, K., Clarke, J., Horn, M., Richards, M., Graham, H., Eisner, D. and Trafford, A.: Characterization of an Extensive Transverse Tubular Network in Sheep Atrial Myocytes and its Depletion in Heart Failure., Circ Heart Fail 2, 482-9, 2009
Protease Type XIV: 0.006%
綿羊綿羊CardiomyocytesCollagenase Type 2: 100-200 u/mlSee referenceDhamoon, A, Pandit, S., Sarmast, F., Parisian, K., Guha, P., Li, Y., Bagwe, S., Taffet, S. and Anumonwo, J.: Unique Kir2.x Properties Determine Regional and Species Differences in the Cardiac Inward Rectifier K+ Current.,Circ Res 94, 1332-9, 2004
Aortic smooth muscle and intersitial cellsCollagenase: 600 u/mlPBSButcher, J. and Nerem, R.: Porcine Aortic Valve Interstitial Cells in Three-Dimensional Culture: Comparison of Phenotype with Aortic Smooth Muscle Cells., J Heart Valve Dis 13, 478, 2004
Coronary vascular smooth muscle (PCVSMCs)Trypsin: 0.037%HEPESChrist, M., Gunther, A., Heck, M., Schmidt, B., Falkenstein, E., and Wehling, M.: Aldosterone, Not Estradiol, Is the Physiological Agonist for Rapid Increases in cAMP in Vascular Smooth Muscle Cells, Circulation 99, 1485, 1999
Miniature swine, adult, female, 25-40 kg, familiarized with treadmill exerciseCoronary smooth muscleTrypsin: 0.1%Krebs bicarbonate solutionBowles, D., Hu, Q., Laughlin, M., and Sturek, M.: Exercise Training Increases L-type Calcium Current Density in Coronary Smooth Muscle, Am J Physiol 275 (44), H2159, 1998
兔, New Zealand, 2.3-2.9 kgCardiomyocytesCollagenase Type 1: 0.05%Krebs-HenseleitFarkas, A., Acsai, K., Nagy, N., Toth, A., Fulop, F., Seprenyi, G., Birinyi, P., Nanasi, P., Forster, T., Csanady, M., Papp, J., Varrro, A. and Farkas, A.:Na(+)/Ca(2+) Exchanger Inhibition Exerts a Positive Inotropic Effect in the Rat Heart, but Fails to Influence the Contractility of the Rabbit Heart., Br J Pharmacol 154, 93, 2008
Hyaluronidase: see reference
Protease: see reference
Smooth muscle cellsCollagenase Type 2: 300 u/mlF10 HamVerheye, S., Martinet, W., Kockx, M., Knaapen, M., Salu, K., Timmermans, J., Ellis, J., Kilpatrick, D., DeMayer, G.: Selective Clearance of Macrophages in Atherosclerotic Plaques by Autophagy, J Am Coll Cardiol49, 706, 2007
Elastase: 5 u/ml
兔, New Zealand, 2 kgCardiomyocytes and fibroblastsCollagenase: 0.06%Medium 199Driesen Ronald B, Dispersyn Gerrit D, Verheyen Fons K, van den Eijnde Stefan M, Hofstra Leo, Thone Fred, Dijkstra Petra, Debie Wiel, Borgers Marcel, Ramaekers Frans C S: Partial cell fusion: a newly recognized type of communication between dedifferentiating cardiomyocytes and fibroblasts,Cardiovasc Res 68, 37-46, 2005
兔, New Zealand White, 2-3 kgMyocytesCollagenase Type 2: 0.1%HEPESSpitzer, K., Ershler, P., Skolnick, R., and Vaughan-Jones, R.: Generation of Intracellular pH Gradients in Single Cardiac Myocytes with a Microperfusion System, Am J Physiol 278, H1371, 2000
兔, New Zealand, white, either sexMyocytesCollagenase Type 2: 0.05%EGTA-KBSedarat, F., Xu, L., Moore, E., and Tibbits, G.: Colocalization of Dihydropyridine and Ryanodine Receptors in Neonate Rabbit Heart Using Confocal Microscopy, Am J Physiol Heart Circ Physiol 279, H202, 2000
兔, adult 1.5-2.5 kgTricuspid valve cellsCollagenase: 0.8 mg/mlHEPES Tyrode solutionAnumonwo JM, Delmar M, and Jalife J.: Electrophysiology of single heart cells from the rabbit tricuspid valve, J Physiol 425, 145, 1990
兔, New Zealand, white, male, 2-3 kgCardiomyocytesHyaluronidase: 0.5%Eagle's MEMBuxton, I., and Brunton, L.: Compartments of Cyclic AMP and Protein Kinase in Mammalian Cardiomyocytes, J Biol Chem 258 (17), 10233, 1983
兔, adult, 2 kgMyocytesHyaluronidase: 0.007%Kreb's RingerDani, A., Cittadini, A., Flamini, G., Festuccia, G., and Terranova, T.:Preparation and Some Properties of Isolated Beating Myocytes from Adult Rabbit Heart, J Mol Cell Cardiol 9, 777, 1977
大鼠大鼠, 1-2 dayCardiomyocytesNCIS kit: per instructionsMEMXie, L., Pi, X., Townley-Tilson, WH., Li, N., Wehrens, X., Entman, M., Taffet, G., Mishra, A., Peng, J., Schisler, J., Meissner, G. and Patterson, C.: PHD2/3-dependent Hydroxylation Tunes Cardiac Response to 尾-adrenergic Stress via Phospholamban., J Clin Invest 125, 2759-71, 2015
大鼠大鼠, Wistar, 250-350gVentricular myocytesCollagenase Type 2: 251 u/mlTyrode solutionCalderon-Sachez, E., Domi璶guez-Rodriguez, A., Lopez-Haldon, J., Jimenez-Navarro, M., Gomez, A., Smani, T. and Ordonez, A.:Cardioprotective Effect of Ranolazine in the Process of Ischemia-reperfusion in Adult Rat Cardiomyocytes., Rev Esp Cardiol (Engl Ed) , , 2015
大鼠大鼠, SD, 8-10 weekVentricular myocytesCollagenase: 0.08%Tyrode's SolutionLee, J., Ha, J. and Leem, C.: A Novel Nicotinamide Adenine Dinucleotide Correction Method for Mitochondrial Ca(2+) Measurement with FURA-2-FF in Single Permeabilized Ventricular Myocytes of Rat., Korean J Physiol Pharmacol 19, 373-82, 2015
Pronase: 0.004%
大鼠大鼠, SD, 1 dayCardiomyocytesTrypsin: 0.025%HBSSClark, A. and Naya, F.: MicroRNAs in the Myocyte Enhancer Factor 2 (MEF2)-regulated Gtl2-Dio3 Noncoding RNA Locus Promote Cardiomyocyte Proliferation by Targeting the Transcriptional Coactivator Cited2., J Biol Chem 290, 23162-72, 2015
Collagenase Type 2: 1.0%
大鼠大鼠, SD, 2-3 dayCardiomyocytesCollagenase Type 2: 0.05%DMEMFang, R, Qiao, S., Liu, Y., Meng, Q., Chen, X., Song, B., Hou, X. and Tian, W.: Sustained Co-Delivery of BIO and IGF-1 by a Novel Hybrid Hydrogel System to Stimulate Endogenous Cardiac Repair in Myocardial Infarcted Rat Hearts., Int J Nanomedicine 10, 4691-703, 2015
大鼠大鼠, SDValve Interstitial cellsCollagenase Type 2: 600 u/mlDMEMHorne, T., VandeKopple, M., Sauls, K., Koenig, S., Anstine, L., Garg, V., Norris, R. and Lincoln, J.: Dynamic Heterogeneity of the Heart Valve Interstitial Cell Population in Mitral Valve Health and Disease, J Cardiovasc Dev Dis 2, 214, 2015
大鼠Wistar, 200 gVentricular myocytesCollagenase Type 2: 95 u/mlHEPESMiller, L., Greensmith, D., Sankaranarayanan, R., O'Neill, S. and Eisner D.:The Effect of 2,5-di-(tert-butyl)-1,4-benzohydroquinone (TBQ) on Intracellular Ca2+ Handling in Rat Ventricular Myocytes., Cell Calcium 58, 208-14, 2015
Protease: 0.1 u/ml
大鼠大鼠, SD, 3-4 monthCardiomyocytesCollagenase Type 2: 0.1%DMEMLiu, L., Wang, Y., Cao, Z., Wang, M., Liu, X., Gao, T., Hu, Q., Yuan, W. and Lin, L.: Up-regulated TLR4 in Cardiomyocytes Exacerbates Heart Failure after Long-Term Myocardial Infarction., J Cell Mol Med 19, 2728, 2015
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsHBSSSong, D., Lee, K., Ryu, J., Jeon, H. and Kim, D.: The Molecular Interaction of Heart LIM Protein (HLP) with RyR2 and Caveolin-3 is Essential for Ca(2+)-Induced Ca(2+) Release in the Heart., Biochem Biophys Res Commun 463, 975-81, 2015
大鼠大鼠, neonatalCardiomyocytesTrypsin: 0.06%HBSSZhang, B., Montgomery, M., Davenport-Huyer, L., Korolj, A. and Radisic, M.: Platform Technology for Scalable Assembly of Instantaneously Functional Mosaic Tissues., Sci. Adv. 1, e1500423, 2015
Collagenase Type 2: 220 u/ml
大鼠大鼠CardiomyocytesCollagenase Type 2: 150 u/mlMEMHennessey, J., Wei, E. and Pitt, G.: Fibroblast Growth Factor Homologous Factors Modulate Cardiac Calcium Channels., Circ Res 113, 381-8, 2013
大鼠大鼠, Wistar, 175-200gCardiomyocytesCollagenase Type 2: 150 u/mlsee referenceEckerle, L., Felix, S. and Herda, L.: Measurement of Antibody Effects on Cellular Function of Isolated Cardiomyocytes., J Vis Exp 73, e4237, 2013
大鼠大鼠, SD, 1-2 dayCardiomyocytesTrypsin: 0.06%DMEMZhang, B., Green, J., Murthy, S. and Radisic, M.: Label-Free Enrichment of Functional Cardiomyocytes using Microfluidic Deterministic Lateral Flow Displacement., PLoS ONE 7, e37619, 2012
Collagenase Type 2: 220 u/ml
大鼠大鼠, adultVentricular myocytesCollagenase Type 2: 0.2%Medium 199Karakikes, I., Hadri, L., Rapti, K., Ladage, D., Ishikawa, K., Tilemann, L., Yi, G., Morel, C., Gwathmey, J., Zsebo, K., Weber, T., Kawase, Y. and Hajjar, R.: Concomitant Intravenous Nitroglycerin with Intracoronary Delivery of AAV1.SERCA2a Enhances Gene Transfer in Porcine Hearts., Mol Therapy 20, 565, 2012
大鼠大鼠, 3 dayCardiomyocytesCollagenase Type 2: 300 u/mlPBSYe, K., Sullivan, K. and Black, L.: Encapsulation of Cardiomyocytes in a Fibrin Hydrogel for Cardiac Tissue Engineering., J Vis Exp 55, 3251, 2011
大鼠大鼠, SD, maleCardiomyocytesCollagenase Type 2: 0.056%KrebsMellor, K., Bell, J., Wendt, I., Davidoff, A., Ritchie, R. and Delbridge, L.:Fructose Modulates Cardiomyocyte Excitation-Contraction Coupling and Ca Handling In Vitro., PLoS ONE 6, e25204, 2011
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Dittami, G., Rajguru, S., Lasher, R., Hitchcock, R. and Rabbitt, R.:Intracellular Calcium Transients Evoked by Pulsed Infrared Radiation in Neonatal Cardiomyocytes., J Physiol 589, 1295-306, 2011
大鼠大鼠, neonatalCardiomyocytesTrypsin: 0.07%DMEM/M199Guan, X., Delo, D., Atala, A. and Soker, S.: In Vitro Cardiomyogenic Potential of Human Amniotic Fluid Stem Cells., J Tissue Eng Regen Med 5, 220, 2011
Collagenase Type 2: 0.1%
大鼠大鼠, adultCardiomyocytesCollagenase Type 2: 0.025%see referenceLouch, W., Sheehan, K. and Wolska, B.: Methods in Cardiomyocyte Isolation, Culture, and Gene Transfer., J Mol Cell Cardiol 51, 288-98, 2011
大鼠大鼠, Wistar, maleCardiomyocytesNCIS kit: per instructionsL-15Brinckmann, M., Kaschina, E., Altarche-Xifro, W., Curato, C., Timm, M., Grzesiak, A., Dong, J., Kappert, K., Kintscher, U., Unger, T. and Li, J.:Estrogen Receptor Alpha Supports Cardiomyocytes Indirectly Through Post-Infarct Cardiac c-kit+ Cells., J Mol Cell Cardiol 47, 66, 2009
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Smith, M., Huang, Y. and Deshmukh, M.: Skeletal Muscle Differentiation Evokes Endogenous XIAP to Restrict the Apoptotic Pathway., PLoS ONE 4, e5097, 2009
大鼠大鼠, adultMyocytesCollagenase Type 2: 0.05%Media 199Xu, X. and Colecraft, H.: Primary Culture of Adult Rat Heart Myocytes., J Vis Exp 28, 1308, 2009
Protease XIV: 0.02%
大鼠大鼠, SD, 200-250 gCardiomyocytesCollagenase Type 1: 0.05%Krebs-HenseleitFarkas, A., Acsai, K., Nagy, N., Toth, A., Fulop, F., Seprenyi, G., Birinyi, P., Nanasi, P., Forster, T., Csanady, M., Papp, J., Varrro, A. and Farkas, A.:Na(+)/Ca(2+) Exchanger Inhibition Exerts a Positive Inotropic Effect in the Rat Heart, but Fails to Influence the Contractility of the Rabbit Heart., Br J Pharmacol 154, 93, 2008
Hyaluronidase: see reference
Protease: see reference
大鼠大鼠, SD, 1-2 dayCardiomyocytesNCIS kit: per instructionsL-15Kim, M., Oh, J., Sakata, S., Liang, I., Park, W., Hajjar, R. and Lebeche, D.:Role of Resistin in Cardiac Contractility and Hypertrophy., J Mol Cell Cardiol 45, 270, 2008
大鼠大鼠, SD, 2 dayCardiomyocytesCollagenase Type 2: 0.1%DMEM/F-12Jang, J., Ku, S., Kim, J., Choi, K., Kim, Y., Kim, H., Oh, S., Lee, E., Cho, H., Song, Y., Lee, S., Lee, S., Suh, C., Kim, S., Moon, S. and Choi, Y.: Notch Inhibition Promotes Human Embryonic Stem Cell-Derived Cardiac Mesoderm Differentiation., Stem Cells 26, 2782, 2008
大鼠大鼠, SD, male, 225-250gCardiomyocytesCollagenase Type 2: 0.1%J-MEMKubli, D., Quinsay, M., Huang, C., Lee, Y., Gustafsson, A.: Bnip3 Functions as a Mitochondrial Sensor of Oxidative Stress During Myocardial Ischemia and Reperfusion., Am J Physiol Heart Circ Physiol 295, H2025, 2008
大鼠大鼠, 2-3 dayCardiomyocytesCollagenase Type 2: 0.05%DMEMLaFramboise, W., Scalise, D., Stoodley, P., Graner, S., Guthrie, R., Magovern, J and Becich, M.: Cardiac Fibroblasts Influence Cardiomyocyte Phenotype in Vitro., Am J Physiol Cell Physiol 292, C1799-808, 2007
Pancreatin: 0.1%
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Robinet, A., Millart, H., Oszust, F., Hornebeck, W. and Bellon, G.: Binding of Elastin Peptides to S-Gal Protects the Heart Against Ischemia/Reperfusion Injury by Triggering the RISK Pathway., FASEB J 21, 1968, 2007
大鼠大鼠, SD, 300-350 gCardiomyocytesCollagenase Type 2: 0.05%DMEMNiederbichler Andreas D, Hoesel Laszlo M, Westfall Margaret V, Gao Hongwei, Ipaktchi Kyros R, Sun Lei, Zetoune Firas S, Su Grace L, Arbabi Saman, Sarma J Vidya, Wang StewartC, Hemmila MarkR, Ward PeterA:An essential role for complement C in the pathogenesis of septic cardiac dysfunction, J Exp Med 203, 53-61, 2006
Hyaluronidase: 0.02%
大鼠大鼠, SD, maleVentricular myocytesCollagenase: 0.13%DMEMStagg Mark A, Coppen Steven R, Suzuki Ken, Varela-Carver Anabel, Lee Joon, Brand Nigel J, Fukushima Satsuki, Yacoub Magdi H, Terracciano Cesare M: Evaluation of frequency, type, and function of gap junctions between skeletal myoblasts overexpressing connexin43 and cardiomyocytes: relevance to cell transplantation, FASEB J 20, 744-6, 2006
Hyaluronidase: 0.06%
大鼠大鼠, SD, 200-250 gVentricular myocytes and mesenteric arterial SMCCollagenase Type 2: 0.06%Krebs-RingerLu Tong, Ye Dan, Wang Xiaoli, Seubert John M, Graves Joan P, Bradbury J Alyce, Zeldin Darryl C, Lee Hon-Chi: Cardiac and vascular KATP channels in rats are activated by endogenous epoxyeicosatrienoic acids through different mechanisms, J Physiol 575, 627-44, 2006
Papain: 0.175%
大鼠大鼠, Wistar, neonatalCardiomyocytesNCIS kit: per instructionsL-15Butler, T., Au, C., Yang, B., Egan, J., Tan, Y., Hardeman, E., North, K., Verkman, A. and Winlaw, D.: Cardiac Aquaporin Expression in Humans, Rats, and Mice., Am J Physiol Heart Circ Physiol 291, H705, 2006
大鼠大鼠, 1-2 dayCardiomyocytesNCIS kit: per instructionsDMEM/F-12Pedram, A., Razandi, M., Aitkenhead, M., and Levin, E: Estrogen Inhibits Cardiomyocyte Hypertrophy in vitro. Antagonism of Calcineurin-related Hypertrophy through Induction of MCIP1, J Biol Chem 280, 26339, 2005
大鼠大鼠, SD, male, 250-300 gCardiomyocytesCollagenase Type 2: 250 u/mlCardioplegic solutionHead, B., Pa, H., Roth, D., Lai, N., Niesman, I., Farquhar, M. and Insel, P.: G-Protein-Coupled Receptor Signaling Components Localize in Both Sarcolemmal and Intracellular Caveolin-3-Associated Microdomains in Adult Cardiac Myocytes., J Biol Chem 280, 31036-44, 2005
大鼠大鼠, Wistar, 1-3 dayCardiomyocytesNCIS kit: per instructionsL-15Castillo, A., Ruzmetov, N., Harvey, K., Stillwell, W., Zaloga, G. and Siddiqui, R.: Docosahexaenoic Acid Inhibits Protein Kinase C Translocation/Activation and Cardiac Hypertrophy in Rat Cardiomyocytes., J Mol Genet Med 1, 18, 2005
大鼠大鼠, SD, 2-4 dayCardiomyocytes, fibroblastsTrypsin: 0.1%Medium 199Entcheva, E., Bien, H, Yin L., Chung, C., Farrell, M., and Kostov, Y.:Functional Cardiac Cell Constructs on Cellulose-based Scaffolding,Biomaterials 25(26), 5753, 2004
Collagenase: 0.1%
大鼠大鼠, SD, 1-2 dayCardiomyocytesNCIS kit: per instructionsL-15Muller-Bore, B., Cascio, W., Anderson, P., Snowwaert, J., Frye, J., Desai, N., Esch, G., Brackham, J., Bagnell, C., Coleman, W., Grisham, J., and Malouf, N.: Adult-derived Liver Stem Cells Acquire a Cardiomyocyte Structural and Functional Phenotype ex vivo, Am J Pathol 165, 135, 2004
大鼠大鼠,1-2 dayCardiomyocytesNCIS kit: per instructionsDMEMNatarajan AR, Rong Q, Katchman AN, and Ebert SN.: Intrinsic cardiac catecholamines help maintain beating activity in neonatal rat cardiomyocyte cultures, Pediatr Res 56, 411, 2004
大鼠大鼠, 1 dayCardiomyocytesNCIS kit: per instructionsF-10Chen, Z., Ge, Y., and Kang J.: Down-regulation of the M6P/IGF-II Receptor Increases Cell Proliferation and Reduces Apoptosis in Neonatal Rat Cardiac Myocytes, BMC Cell Biol 5, 15, 2004
大鼠大鼠, neonatalCardiac myocytesCollagenase Type 2: 0.5%HBSSChen Hsiao-Huei, Mullett Steven J, Stewart Alexandre F R: Vgl-4, a novel member of the vestigial-like family of transcription cofactors, regulates alpha1-adrenergic activation of gene expression in cardiac myocytes, J Biol Chem 279, 30800-6, 2004
大鼠大鼠, male, 250-300 gCardiomyocytesCollagenase Type 1: 0.03%HBSSHunton DaciaL, Zou LuYun, Pang Yi, Marchase Richard B: Adult rat cardiomyocytes exhibit capacitative calcium entry, Am J Physiol Heart Circ Physiol 286, H1124-32, 2004
Protease: 0.01%
大鼠大鼠, neonatalCardiomyocytesCollagenase Type 2: 0.1%DMEMTamamori-Adachi, M., Hayashida, K., Nobori, K., Omizu, C., Yamada, K., Sakamoto, N., Kamura, T., Fukuda, K., Ogawa, S., Nakayama, K. and Kitajima, S.: Down-Regulation of p27 Promotes Cell Proliferation of Rat Neonatal Cardiomyocytes Induced by Nuclear Expression of Cyclin D1 and CDK4, J Biol Chem 2279, 50429, 2004
大鼠大鼠, fetal or 3-dayCardiomyocytesCollagenase Type 2: 0.1%HEPESTamamori-Adachi Mimi, Ito Hiroshi, Sumrejkanchanakij Piyamas, Adachi Susumu, Hiroe Michiaki, Shimizu Masato, Kawauchi Junya, Sunamori Makoto, Marumo Fumiaki, Kitajima Shigetaka, Ikeda Masa-Aki: Critical role of cyclin D1 nuclear import in cardiomyocyte proliferation, Circ Res 92, e12-9, 2003
大鼠大鼠, adult, 220-280gCardiomyocytesCollagenase Type 2: 178 u/mlKrebs-Henseleit, CFGordon Jennifer M, Dusting Gregory J, Woodman Owen L, Ritchie Rebecca H: Cardioprotective action of CRF peptide urocortin against simulated ischemia in adult rat cardiomyocytes, Am J Physiol Heart Circ Physiol 284, H330-6, 2003
Hyaluronidase: 0.01%
大鼠大鼠, 50 gVentricular myocytesCollagenase: 223 u/mlMedium 199Aberle II NS, and Ren J.: Experimental Assessment of the Role of Acetaldehyde in Alcoholic Cardiomyopathy, Biol Proced Online 5, 1, 2003
Hyaluronidase: 0.01%
Trypsin: 0.002%
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Li TS, Ito H, Kajiwara K, and Hamano K.: Long-term survival of xenografted neonatal cardiomyocytes by adenovirus-mediated CTLA4-Ig expression and CD40 blockade, Circulation 108, 1760, 2003
大鼠大鼠, SD, 200-300 gCardiomyocytesCollagenase Type 2: 0.05-0.1%Joklik's MEMSwift Luther, McHowat Jane, Sarvazyan Narine: Inhibition of membrane-associated calcium-independent phospholipase A2 as a potential culprit of anthracycline cardiotoxicity, Cancer Res 63, 5992-8, 2003
大鼠大鼠, SD, 175-200gCardiomyocytesCollagenase Type 2: 0.05%Krebs-RingerTardif Annie, Julien Nathalie, Chiasson Jean-Louis, Coderre Lise:Stimulation of glucose uptake by chronic vanadate pretreatment in cardiomyocytes requires PI 3-kinase and p38 MAPK activation, Am J Physiol/Endo 284, E1055-64, 2003
Deoxyribonuclease I: 0.02%
大鼠大鼠, SD, pregnant femalePeritoneal mastHyaluronidase: 100 u/mlDMEM (see reference)DeAlmeida, A., Mustin, D., Forman, M., Brower, G., Janicki, J., and Carver, W.: Effects of mast cells on the behavior of isolated heart fibroblasts: modulation of collagen remodeling and gene expression, J Cell Physiol 191, 51, 2002
大鼠大鼠, neonatal, adult, 17 dayVentricular cardiomyocytesCollagenase Type 2: 0.7-1%DMEMLam ML, Bartoli M, and Claycomb WC.: The 21-day postnatal rat ventricular cardiac muscle cell in culture as an experimental model to study adult cardiomyocyte gene expression, Mol Cell Biochem 229, 51, 2002
大鼠大鼠, 1 dayCardiomyocytesNCIS kit: per instructionsL-15Yang Y, Liao H, Ke Q, Cai J, Xiao YF, and Morgan JP: Enhancement of nitric oxide production by methylecgonidine in cultured neonatal rat cardiomyocytes, Br J Pharmacol 135, 188, 2002
大鼠大鼠, female, 250-300 gVentricular myocytesCollagenase Type 2: 0.1%Ringer solutionLiu Qinghang, Hofmann Polly A: Antiadrenergic effects of adenosine A(1) receptor-mediated protein phosphatase 2a activation in the heart, Am J Physiol Heart Circ Physiol 283, H1314-21, 2002
大鼠大鼠, Wistar, 1 dayCardiomyocytesCollagenase Type 2: 80u/mlM199Shimizu Tatsuya, Yamato Masayuki, Isoi Yuki, Akutsu Takumitsu, Setomaru Takeshi, Abe Kazuhiko, Kikuchi Akihiko, Umezu Mitsuo, Okano Teruo: Fabrication of pulsatile cardiac tissue grafts using a novel 3-dimensional cell sheet manipulation technique and temperature-responsive cell culture surfaces, Circ Res 90, e40, 2002
大鼠大鼠, male, 6-12 months oldCardiac myocytesProtease: 0.55 u/mlTyrode's solutionKitta, K., Day, R., Ikeda, T., and Suzuki, Y.: Hepatocyte Growth Factor Protects Cardiac Myocytes Against Oxidative Stress-Induced Apoptosis, Free Radic Biol Med 31 (7), 902, 2001
大鼠大鼠, Harlan SD, pupCardiomyocytesTrypsin: DMEM/Ham's F-12Schwartzbauer, G., and Robbins, J.: The Tumor Suppressor Gene PTEN Can Regulate Cardiac Hypertrophy and Survival, J Biol Chem 276, 35786, 2001
大鼠大鼠, SD, newbornCardiac myocytesNCIS kit: per instructionsL-15Rahman A, Alam M, Rao S, Cai L, Clark LT, Shafiq S, and Siddiqui MA:Differential effects of doxorubicin on atrial natriuretic peptide expression in vivo and in vitro, Biol Res 34(3-4), 195, 2001
大鼠大鼠, adultVentricular myocytesCollagenase: 223 u/mlTyrode solutionRen J, and Wold LE.: Measurement of Cardiac Mechanical Function in Isolated Ventricular Myocytes from Rats and Mice by Computerized Video-Based Imaging, Biol Proced Online 3, 43, 2001
Hyaluronidase: 0.01%
Trypsin: 0.002%
大鼠大鼠, SD, maleCardiomyocytesCollagenase Type 2: 140 u/mlKrebs-HenseleitDai L, Brookes PS, Darley-Usmar VM, and Anderson PG.: Bioenergetics in cardiac hypertrophy: mitochondrial respiration as a pathological target of NO, Am J Physiol/Heart 281, H2261-H2269, 2001
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsDMEM/M199Guo K, Searfoss G, Krolikowski D, Pagnoni M, Franks C, Clark K, Yu KT, Jaye M, Ivashchenko Y: Hypoxia induces the expression of the pro-apoptotic gene BNIP3, Cell Death Differ 8, 367-76, 2001
大鼠大鼠, Wistar, femaleVentricular myocytesCollagenase Type 2: 0.1%Ringer solutionPyle, W, Lester, J, and Hofmann, P.: Effects of Kappa-opioid receptor activation of myocardium, Am J Physiol Heart Circ Physiol 281, H669, 2001
大鼠大鼠, SD, 1-2 dayCardiomyocytesTrypsin: 0.01%HBSSArutunyan A, Webster D, Swift L, Sarvazyan N: Localized injury in cardiomyocyte network: a new experimental model of ischemia-reperfusion arrhythmias, Am J Physiol Heart Circ Physiol 280, H1905-15, 2001
Collagenase Type 2: 0.08%
大鼠大鼠, newbornAtrialTrypsin: 0.06%DMEMKim, D., and Pleumsamran, A.: Cytoplasmic Unsaturated Free Fatty Acids Inhibit ATP-Dependent Gating of the G Protein-gated K+ Channel, J Gen Physiol 115, 287, 2000
大鼠大鼠, 1-2 dayCardiomyocytesCollagenase Type 1: 84 u/mlM199 mediumBueno OF, De Windt LJ, Tymitz KM, Witt SA, Kimball TR, Klevitsky R, Hewett TE, Jones SP, Lefer DJ, Peng CF, Kitsis RN, and Molkentin JD.: The MEK1-ERK1/2 signaling pathway promotes compensated cardiac hypertrophy in transgenic mice, EMBO J 19(23), 6341, 2000
Pancreatin: 0.05%
大鼠大鼠, Winstar, 1-2 dayVentricular myocytesCollagenase Type 2: 0.08%DMEMMaki T, Horio T, Yoshihara F, Suga S, Takeo S, Matsuo H, and Kangawa K: Effect of neutral endopeptidase inhibitor on endogenous atrial natriuretic peptide as a paracrine factor in cultured cardiac fibroblasts, Br J Pharmacol 131(6), 1204, 2000
大鼠大鼠, SD, adult femaleHeart microvascular cellsCLSPA: 250u/mlL-15Puri S, Bansal DD, Uskokovic MR, and MacGregor RR.: Induction of tissue plasminogen activator secretion from rat heart microvascular cells by fM 1,25(OH)(2)D(3), Am J Physiol/Endo 278(2), E293, 2000
Papain: 5 u/ml
Elastase: 0.8 u/ml
大鼠大鼠, SD, male, 250-300gCardiomyocytesCollagenase Type 2: 0.06%M199Sun L, Chang J, Kirchhoff SR, and Knowlton AA.: Activation of HSF and selective increase in heat-shock proteins by acute dexamethasone treatment, Am J Physiol Heart Circ Physiol 278(4), H1091, 2000
大鼠大鼠, wistar, 2 dayCardiomyocytesNCIS kit: per instructionsDMEM-F12Cowan DB, Poutias DN, Del Nido PJ, and McGowan FX Jr: CD14-independent activation of cardiomyocyte signal transduction by bacterial endotoxin, Am J Physiol Heart Circ Physiol 279, H619, 2000
大鼠大鼠, neonatalCardiomyocytesNCIS kit: per instructionsL-15Webster DR, and Patrick DL.: Beating rate of isolated neonatal cardiomyocytes is regulated by the stable microtubule subset, Am J Physiol Heart Circ Physiol 278, H1653, 2000
大鼠大鼠, Lewis, adult male, 300-350-gAorta, smooth muscleTrypsin: 0.025%Medium 199 with 20% FBS mediumKim, B., Nikolovski, J., Bonadio, J., Smiley, E., and Mooney, D.: Engineered Smooth Muscle Tissues: Regulating Cell Phenotype with the Scaffold, Exp Cell Res 251, 318, 1999
大鼠大鼠, 10-15 newborn, 1-2 days oldAtrialTrypsin: 0.03%HBSS or EBSSNemec, J., Wickman, K., and Clapham, D.: GBy Binding Increases the Open Time of IKACh: Kinetic Evidence for Multiple GBy Binding Sites, Biophys J 76, 246, 1999
大鼠大鼠, 1-3 dayNeonatal rat cardiomyocytesCollagenase Type 2: 80u/mlDMEM/F12Adderley SR, and Fitzgerald DJ.: Oxidative damage of cardiomyocytes is limited by extracellular regulated kinases 1/2-mediated induction of cyclooxygenase-2, J Biol Chem 274, 5038, 1999
Pancreatin: 0.06%
大鼠大鼠, 1-2 dayCardiomyocytesPancreatin: 0.06%DMEMReinecke H, Zhang M, Bartosek T, and Murry CE.: Survival, integration, and differentiation of cardiomyocyte grafts: a study in normal and injured rat hearts, Circulation 100(2), 193, 1999
Collagenase Type 2: 95 u/ml
大鼠大鼠, adult and neonatalCardioomyocytesCollagenase Type 4: 0.12%-adultDMEM/medium 199Richards SM, Jaconi ME, Vassort G, and Puceat M.: A spliced variant of AE1 gene encodes a truncated form of Band 3 in heart: the predominant anion exchanger in ventricular myocytes, J Cell Sci 112, 1519, 1999
Collagenase Type 2: 0.05%-neonatal
大鼠大鼠, Wistar, maleCardiomyocytesCollagenase Type 2: 228 u/mlJoklik MEMSambandam N, Abrahani MA, St Pierre E, Al-Atar O, Cam MC, and Rodrigues B.: Localization of lipoprotein lipase in the diabetic heart: regulation by acute changes in insulin, Arterioscler Thromb Vasc Biol 19(6), 1526, 1999
大鼠大鼠, 1-2 dayCardiomyocytesNCIS kit: per instructionsRPMIMatsui, T., Li, Ling, Del Monte, F., Fukui, Y., Franke, T., Hajjar, R., Rosenzweig, A.: Adenoviral gene transfer of activated phosphatidylinositol 3'-kinase and Akt inhibits apoptosis of hypoxic cardiomyocytes in vitro,Circulation 100, 2373-9, 1999
大鼠大鼠Thoracic aorta, smooth muscleElastase: 0.02%(see reference)Zilberman, A., Dave, V., Miano, J., Olson, E., and Periasamy, M.:Evolutionary Conserved Promoter Region Containing CArg-Like Elements is Crucial for Smooth Muscle Myosin Heavy Chain Gene Expression, Circ Res 82, 566, 1998
大鼠大鼠, SD, 1 dayCardiomyocytesNCIS kit: per instructionsDMEM/F-12Wagner DR, Combes A, McTiernan C, Sanders VJ, Lemster B, Feldman AM: Adenosine inhibits lipopolysaccharide-induced cardiac expression of tumor necrosis factor-alpha, Circ Res 82, 47-56, 1998
大鼠大鼠, SD, 300-350 gCardiomyocytesCollagenase Type 2: 0.04%see referenceYu L, Netticadan T, Xu YJ, Panagia V, Dhalla NS: Mechanisms of lysophosphatidylcholine-induced increase in intracellular calcium in rat cardiomyocytes, J Pharmacol Exp Ther 286, 1-8, 1998
大鼠大鼠, SD, female, 200gVentricular cardiac myocytesCollagenase Type 2: 0.5%Krebs-Henseleit bufferWestfall M V, Rust E M, Metzger J M: Slow skeletal troponin I gene transfer, expression, and myofilament incorporation enhances adult cardiac myocyte contractile function, Proc Natl Acad Sci U S A 94, 5444-9, 1997
Hyaluronidase: 0.2%
大鼠大鼠, Wistar, 1 dayCardiomyoctesCollagenase Type 2: 80 u/mlHBSSKinugawa, K., Kohmoto, O., Yao, A., Serizawa, T., and Takahashki, T.:Cardiac inducible nitric oxide synthase negatively modulates myocardial function in cultured rat myocytes, Am J Physiol 272, H35-47, 1997
大鼠大鼠, neonatal, 5-dayCardiomyocytesTrypsin: 0.05%DMEMVanwinkle, W., Snuggs, M., and Buja, L.: Cardiogel: A Biosynthetic Extracellular Matrix for Cardiomyocyte Culture, In Vitro Cell Dev Biol Anim 32, 478, 1996
大鼠大鼠, SD, 250-300 gCardiomyocytesCollagenase Type 2: 0.05%Joklik'sSharma VK, Colecraft HM, Wang DX, Levey AI, Grigorenko EV, Yeh HH, and Sheu SS: Molecular and functional identification of m1 muscarinic acetylcholine receptors in rat ventricular myocytes, Circ Res 79(1), 86, 1996
大鼠大鼠, SD, fetal and neonatalCardiomyocytesCollagenase: 0.1%PBSLi RK, Mickle DA, Weisel RD, Zhang J, Mohabeer MK: In vivo survival and function of transplanted rat cardiomyocytes, Circ Res 78, 283-8, 1996
Trypsin: 0.2%
大鼠SD, SHR or WKYCardiomyocytesNCIS kit: per instructionsHBSSMacGregor, R., Klein, R., and Bansal, D.: Secretion of Plasminogen Activator Activity from Neonatal Rat Heart Cells is Regulated by Hormones and Growth Factors, Ann N Y Acad Sci 752, 331, 1995
大鼠大鼠, SD, male, 200-250 gMyocytesCollagenase: 0.07%Joklik's MEMLaughlin, M., Schaeffer, M., and Sturek, M.: Effect of Exercise Training on Intracellular Free Ca2+ Transients in Ventricular Myocytes of Rats, Am J Physiol 73, 1141, 1992
Ventricles
大鼠大鼠, SD, female, 60-90 daysMyocardialCollagenase Type 2: 0.1%Joklik's MEM, Kreb's-Henseleit buffer, CFWelder, A.A., Grant, R., Bradlaw, J., and Acosta, D.: A Primary Culture System of Adult Rat Heart Cells for the Study of Toxicologic Agents, In Vitro Cell Dev Biol 27, 921, 1991
大鼠大鼠, SD, adult, female, 180-220 gCardiomyocytesCollagenase: 0.11%(see reference)Fischer, Y., Rose, H., and Kammermeier. H.: Highly Insulin-Responsive Isolated Rat Heart Muscle Cells Yielded By a Modified Isolation Method, Life Sci 49, 1679, 1991
大鼠大鼠, neonatal, 1-4 daysCardiomyocytesCollagenase Type 2: 0.12%Krebs-Henseleit Buffer, CFMarino, T.A., Walter, R.,Cobb, E., Palasiuk, M., Parsons, T., and Mercer, W.E.: Effects of Norepinephrine on Neonatal Rat Cardiocyte Growth and Differentiation, In Vitro Cell Dev Biol 26, 229, 1990
大鼠大鼠 HeartTrypsin: 0.06%25mM HEPES buffered Locke's solution, CMFTrifaro, J., Tang, R., and Novas, M.: Monolayer Co-Culture of Rat Heart Cells and B綿羊 Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990
(also b綿羊)Adrenal chromaffin 
 Paraneurons
大鼠大鼠, adult, femaleMyocytesCollagenase Type 2: 0.05%Joklik's MEMNag, A.C., Lee, M., and Kosiur, J.R.: Adult Cardiac Muscle Cells in Long-Term Serum-Free Culture:Myofibrillar Organization and Expression of Myosin Heavy Chain Isoforms, In Vitro Cell Dev Biol 26, 464, 1990
大鼠大鼠s, AW, late-gestation pregnant femalesVentricularCollagenase Type 2: 0.05 - 0.08%See referenceEngelmann, G., McTiernan, C., Gerrity, R., and Samarel, A.: Technique 2 (6), 279, 1990
Cardiomyocytes
大鼠大鼠,SD, albino, 10-14 weeksMyocytesCollagenase Type 1: 100 u/ml and 150 u/mlKrebs Ringer bicarbonate bufferBerg, I., Guse, A.H., and Gercken, G.: Carbamoylcholine-Induced Accumulation of Inositol Mono-,Bis-,Tris-, And Tetrakisphosphates In Isolated Cardiac Myocytes From Adult Rats, Biochim Biophys Acta 1010, 100, 1989
大鼠大鼠, SD, adult, female, 175 - 225 gMyocytesCollagenase: 0.1%Joklik's mediumBugaisky, L.B. and Zak, R.: Differentiation of Adult Rat Cardiac Myocytes in Cell Culture, Circ Res 64, 493, 1989
大鼠大鼠, SD, male, 250 - 350 gMyocytes, ventricularCollagenase Type 1: 90 and 100 u/mlJoklik's solution with and without calciumDe Young, M., Giannattasio, B., and Scarpa, A.: Isolation of Calcium-Tolerant Atrial And Ventricular Myocytes From Adult Rat Heart,Methods Enzymology Vol. 173,, , 662, 1989
大鼠大鼠, SD, 2-4 day oldCardiomyocytesTrypsin: 0.1%HBSSToraason, M., Luken, M., Breitenstein, M., Krueger, J., and Biagini, R.:Comparative Toxicity of Allylamine and Acrolein in Cultured Myocytes and Fibroblasts from Neonatal Rat Heart, Toxicology 56, 107, 1989
Fibroblasts
大鼠大鼠, SD, maleAortic smooth muscleTrypsin: 0.05%DMEMCornwell, T., and Lincoln, T.: Regulation of Intracellular Ca2+ Levels in Cultured Vascular Smooth Muscle Cells, J Biol Chem 264 (2), 1146, 1989
大鼠大鼠, adult, 150-200 gMyocytes, atriaCollagenase Type 1: 100 u/mlKreb's Ringer bicarbonate - HEPES bufferMcMahon, K.K.: A Study of Adult Rat Atrial Myocyte Attachment to Extricellular Matrix Components and Long Term Culture,Biology of Isolated Adult Cardiac Myocytes,(Clark, Decker, Borg, Eds), Elsevier Science Publishing Co. Inc., 318, 1988
大鼠SD大鼠MyocytesCollagenase Type 2: 0.1%Kreb's Ringer w/ calciumBuxton, I.L.O. and Doggwiler, K.O.: Alpha1-Adrenergic Receptor Signal Transduction in the Adult Rat Cardiac Myocyte,Biology of Isolated Adult Cardiac Myocytes,(Clark, Decker, Borg, eds), , 298, 1988
大鼠SD大鼠, 雄性, 250-350 克Myocytes, heartHyaluronidase: 0.03%Bicarbonate-buffered mediumKim, D. and Smith, T.W.: Temporal Variation in Contractile State And [Ca++] in Isolated Adult Rat And Guinea Pig Cardiac Myocytes,Biology of Isolated Adult Cardiac Myocytes,(Clark, Decker, Borg, eds), Elsevier Science Publishing Co. Inc., 370, 1988
大鼠Wistar大鼠,  2-4 天neonatesMyoblast, cardiacTrypsin: 0.1%Standardized MediumGrynberg, A., Degois, M., Guenot, L., and Athias, P.: Primary Rat Cardiac Cell Culture: Diet of the Mother Rats as a Determinant Parameter of Cardiomyoblast Production from Neonates, Biol Cell 57, 89, 1986
(see reference)
大鼠大鼠, SD, 懷孕的MyocytesTrypsin: 2.4 u/mlDMEMFreerksen, D., Schroedl, N., and Hartzell, C.: Control of Enzyme Activity Levels by Serum and Hydrocortisone in Neonatal Rat Heart Cells Cultured in Serum-Free Medium, J Cell Physiol 120, 126, 1984
大鼠大鼠, SD, 雌性, 12 周MyocytesCollagenase Type 1: 0.06%Krebs Ringer bicarbonate bufferPiper, H.M., Probst, I., Schwartz, P., Hutter, F.J., and Spieckermann, P.G.:Culturing of Calcium Stable Adult Cardiac Myocytes, J Mol Cell Cardiol 14, 397, 1982
大鼠大鼠, 雄性/雌性, 200-250 克MuscleHyaluronidase: 0.1%Krebs Ringer phosphate buffer, CMFNag, A. and Cheng, M.: Adult Mammalian Cardiac Muscle Cells in Culture,Tissue Cell 13, 515, 1981
大鼠大鼠, 雌性, 成年, 200 - 300 克MyocytesCollagenase Type 2: 0.1%Joklik's MEMClaycomb, W.C., and Palazzo, M.C.: Culture of The Terminally Differentiated Adult Cardiac Muscle Cell: A Light and Scanning Electron Microscope Study,Dev Biol 80, 466, 1980
大鼠SD大鼠, 成年, 雄性, 200-250 克MyocytesHyaluronidase: 0.10%Joklik MEM, CFFrangakis, C., Bahl, J., McDaniel, H., and Bressler, R.: Tolerance to Physiological Calcium by Isolated Mycocytes from the Adult Rat Heart; An Improved Cellular Preparation, Life Sci 27, 815, 1980
大鼠SD大鼠, 雌性, 12 周, 250 克MyocytesCollagenase Type 1: 0.20%Kreb's Ringer bicarbonate bufferPowell, T., Terrar, D.A., and Twist, V.W.: Electrical Properties of Individual Cells Isolated From Adult Rat Ventricular Myocardium, J Physiol 302, 131, 1980
Ventricular myocardium
大鼠大鼠, 10 周MyocytesCollagenase Type 1: 0.1%SalineBishop, S., Oparil, S., Reynolds, R., and Drummond, J.: Regional Myocyte Size in Normotensive and Spontaneously Hypertensive Rats, Hypertension 1 (4), 378, 1979
大鼠大鼠, 雄性, 200-300 克MyocytesHyaluronidase: 82 u/mlKreb's Henseleit bufferFarmer, B., Harris, R., Jolly, W., Hathaway, D., Katzberg, A., Watanabe, A., Whitlow, A., and Besch, H.: Isolation and Characterization of Adult Rat Heart Cells, Arch Biochem Biophys 179, 545, 1977
大鼠大鼠,雌性, 白化變種, 200-250克MyocytesHyaluronidase: 0.1%Kreb's Ringers phosphateNag, A., Fischman, D., Aumont, M., and Zak, R.: Studies of Isolated Adult Rat Heart Cells: The Surface Morphology and the Influence of Extracellular Calcium Ion Concentration on Cellular Viability, Tissue Cell 9 (3), 419, 1977
大鼠大鼠, 成年, 雄性, 150-200克MyocytesCollagenase: 0.1%MEM CFGrosso, D., Frangakis, C., Carlson, E., and Bressler, R.: Isolation and Characterization of Myocytes From the Adult Rat Heart, Prep Biochem 7 (5), 383, 1977
Hyaluronidase: 0.1%
大鼠大鼠, 成年的MyocytesCollagenase: 0.1%Perfusing solutionMoustafa, E., Skomedal, T., Osnes, J., and Oye, I.: Cyclic Amp Formation and Morphology of Myocardial Cells Isolated from Adult Heart: Effect of Ca2+ and Mg2+, Biochim Biophys Acta 421, 411, 1976
大鼠 SD大鼠, 白化變種, 雄性, 180-230 克Heart ventricles, beatingCollagenase: 0.05-0.2%Phosphate bufferGlick, M., Burns, A., and Reddy, W.: Dispersion and Isolation of Beating Cells from Adult Rat Heart, Anal Biochem 61, 32, 1974
大鼠大鼠, 希伯來聯(lián)合國, 3 & 17 個(gè)月, 150-210 克MyocytesTrypsin: 0.05%Versene bufferBierman, E.L., Stein, O., and Stein, Y.: Lipoprotein Uptake and Metabolism by Rat Aortic Smooth Muscle Cells in Tissue Culture, Circ Res 35, 136, 1974
大鼠大鼠, 2-5 天MyocytesTrypsin: 0.01%Saline ASpeicher, D., and McCarl, R.: Pancreatic Enzyme Requirements for the Dissociation of Rat Hearts for Culture, In Vitro 10, 30, 1974
大鼠大鼠, SD, male, 150-200克MyocytesHyaluronidase: 0.05%(see reference)Pretlow II, T., Glick, M., and Reddy, W.: Separation of Beating Cardiac Myocytes from Suspensions of Heart Cells, Am J Pathol 67 (2), 215, 1972
大鼠大鼠, albino, male, 250 克MyocytesHyaluronidase: 0.20%Hank's solution, CFBerry, M., Friend, D., and Scheuer, J.: Morphology and Metabolism of Intact Muscle Cells Isolated from Adult Rat Heart, Circ Res 26, 679, 1970
大鼠大鼠, 成年的, 雄性, 250 克MyocytesTrypsin: 0.1%Saline AVahouny, G.V., Wei, R., Starkweather, R., and Davis, C.: Preparation of Beating Heart Cells From Adult Rats, Science 167, 1616, 1970
大鼠Wistar大鼠,  1-4 天大MyocytesTrypsin: 0.125%Gey's BSSMark, G., and Strasser, F.: Pacemaker Activity and Mitosis in Cultures of Newborn Rat Heart Ventricle Cells, Exp Cell Res 44, 217, 1966
貝類動物太平洋牡蠣HaemocytesTrypsin: L15 mediumLe Duff, R., Lipart, C., and Renault, T.: Primary Culture of Pacific Oyster, Crassostrea gigas, heart cells, J Tiss Cul Meth 16, 67, 1994
貝類動物蚌, 3.0 - 4.5 厘米MyocytesCollagenase Type 1: 2%L15 mediumWen, C., Kou, G., and Chen, S.: C*tion of Cells From the Heart of the Hard Clam, Merretrix lusoria (Roding), J Tiss Cul Meth 15, 123, 1993
Fibroblasts

 

我們公司zui大優(yōu)勢是強(qiáng)大的采購,

1:基本什么都能進(jìn)口,血清,抗體,耗材,還有部分限制進(jìn)口的,

2:貨品全,現(xiàn)經(jīng)營過700多個(gè)品牌,基本所有生物試劑耗材都可以進(jìn)口,特別是冷偏的產(chǎn)品那就更有優(yōu)勢,

3:提供加急服務(wù),一般1-2周到貨,超過時(shí)限加急費(fèi)全免

4:價(jià)格公道,絕大部分價(jià)格有優(yōu)勢,當(dāng)然不能保證100%產(chǎn)品都是,因?yàn)橐馕吨鴽]有服務(wù).

5:良好的信譽(yù),大部分客戶我們提供貨到付款服務(wù),客戶包括清華,北大 交大 復(fù)旦,中山等100多所大學(xué),ROCHE,阿斯利康,國藥 ,fisher等500多家公司

6:我們還是Santa,Advanced Biotechnologies Inc, Worthington Biochemical ,bangs,BBInternational,crystalchem,dianova,FD Neurotechnologies,Inc. Dako,Inc, GePromegaridege, Glycotope Biotechnology GmbH; iduron,Innovative Research of America, Ludger, neuroprobe,Worthington Biochemical , Polysciences,prospecbi, QA-BIO,quickzyme,FULLERLABS,INC,sterlitech;sysy,TriLink BioTechnologies,Inc;worthington-biochem,zyagen等幾十家國外公司授權(quán)代理。
7:我們還是invitrogen,qiagen,Worthington Biochemical am,sigma;Promega,roche,merck, rnd,BD, GE,pierce,BioLegend等*批發(fā),歡迎合作。

 

99热在线观看| 9色91视频| 欧美丁香五月夫妻天| 91色婷婷综合久久中文字幕二区| eeuus五月婷| 久久久月丁香| 婷婷综合成人| 国产偷人爽久久久久久老妇APP| 激情五月天婷婷| 精品99在线| 久久久五月天| 婷婷精品综合| 91五月天| 久久婷婷青青| 亚洲免费看片| 亚洲日日操| 国产成人av在线播放| 秋霞免费视频| 五月天婷综合| 五月天激情国产综合AV| 天堂久久婷婷| 色狠狠色综合久久久绯色aⅴ影视| 色婷婷啪啪| 九九精品热| 影音先锋人妻出差| 久操热线| 丁香社92视频| 蜜臀av无码久久久久久久久| 欧美另类图片| 黄网在线免费| 五月婷婷丁香深深爱| 成年人最刺激的综合网| 91/九色黑人| 色色丁香| 久操人妻| 色综合久久天天综合网| 激情文学五月丁香六月婷婷| 亚洲一区在线播放| 东京热伊人| 婷婷五月天男人影院色色网| 一点色成人网| 亚洲视频在线观看99| 日狠狠| 99re6在线视频精品免费| 成人av中文字幕| 俺去啦综合网| 青青视频在线观看免费2| 伊人99久久| 91九九| 国产做A爰片毛片A片美国| 婷婷久久综合久| 91色五月| 久久视频这里有精品99| www999日韩精品| 五月综合婷婷久久在线| 精品亚洲麻豆1区2区3区| 色五月xxx| 激情综合五月| 色射7856五月天激情四射| 99er免费在线观看| 色色色综合网| 九九九九国产| 欧美大肥婆大肥BBBBB| 色噜噜97视频在线观看| 狠狠干在线| 五月婷婷影视| 99久久精品网| 性欧美大战久久久久久久83| 一级操逼内射在线视频| 亚洲av电影在线| 天天射射夜| 婷婷狠狠干| 久久视频婷婷| 婷婷五月在线视频| 成人国产综合| 日韩一区二区A片免费观看| 九热...av| 日本美女五月天| 中文在线成人| 大地资源中文在线观看免费| 91狠狠综合久久久久久| 九九伊人网| 第四色婷婷五月| 欧美性色A片免费免费观看的| 六月丁香啪啪| 亚洲激情免费视频| 婷婷五月丁香成人网| 青青草深爱激情网| 亚洲在线中文字幕2| 久热久色| 熟妇无码乱子成人精品| 人人视频色| 丁香五月婷综合| 特级毛片AAAAAA| 伊人狠狠丁香婷婷综合尤物| 亚洲天堂久久| 天天日日人| 久久天堂网| 婷婷色激情五月天| 婷婷精品视频| 天天插操| 五月婷婷丁香五月 | 激情婷婷| 五月丁香婷婷啪啪综合网| 欧美激情性做爰免费视频| 激情五月婷婷综合视频| 色婷婷精品视频| 色色国产| 亚洲色爽| 色婷婷综合在线| 久久资源网五月婷| 婷婷五月天首页| 久久综合婷婷五月| 99热精品在线| 人人人va亚洲视频在线| 五月婷精品| 国产乱人偷精品人妻A片| 激情五月天色婷婷综合| 亚洲另类婷婷综合| 极骚大香蕉伊人| 天天天天干| 天天开心天天色| 极品少妇XXXX精品少妇偷拍| 亚洲欧美一区二区三区爱爱动图| 午夜丁香综合婷婷| 91欧美| 婷婷综合色播网| 98色花堂98t.R| 精品亚洲麻豆1区2区3区| 婷婷婷婷婷婷婷婷| 亚洲婷婷丁香五月在线| 久久大香蕉丁香| 天天婷婷色六月| 超碰成人电影| 狠狠色婷婷丁香五月| 级情九色| 婷婷伊人五月| 天堂AV三级| 成年免费大片黄在线观看岛国| 免费在线观看欧美激情xx小视频| 成人婷婷色五月天| 激情啪啪五月天| 久久五月婷综合网| 婷婷五月大香蕉| 最近中文字幕大全免费版在线| 天天综合中文| 99热最新精品| 婷久久久| 色国产在线视频一区| 色婷婷激情四射视频| 九九色综合| 极品嫩草| 97丨九色丨国产丨PORNY| 久久精品4| 日本va欧美va欧美va| 开心五月六月婷婷| 亚洲国产成人AV在线| 99在线视频播放| 99爱这里只有精品| 激情综合五月天| 91免费啪视频| 亚洲天堂婷婷丁香| 熟女人妻视频| 人人爱人人草| 婷婷五月天伊人网| 五月色色色| 五月丁香激情婷婷综合| va中文资源在线观看| 婷婷99狠狠躁天天躁中| 五月天色色色色色| 91热在线观看视频| 九九九九热99超碰| 日逼AV影音先锋男人资源站| 国产精品99久久久久久久女警| 六月丁香中文字幕| 九九在线精品| 久热免费| 成年人最刺激的综合网| 日本高清综合网五月丁香| 《》【无码】想被搞到爽AV应募而来的超M素人 西纯子 10musume-011723-01 | 九色成人AV在线| 欧美成人精品A片免费一区99| 99热综合网| 亚洲bt丁香五月天婷婷激情小说| 婷婷激情丁五月| 立川无码av| 五月丁香六月婷婷亚洲天堂网站| 在线不卡视频| 色婷婷综合网站| 九九自拍网| 4399伦理午夜| 久久久久亚洲AV无码网影音先锋| 无码yw| 综合日本婷婷| 久久久精品免费啪啪国| 3p九色在线| 亚洲综合网在线| 九九综合色| 亚洲中文字幕网| 色99在线观看| 热久久91| 国产精品成人AV在线| 99热在线观看| 久久色五月| 五月天天爽| 激情五月丁香六月| 色5月丁香婷婷| 成年人最刺激的综合网| 九九大香蕉黄色影院| 国产精品久久7777777精品无码| 午夜国产精品AV在线播放| 涩涩婷婷五月| 色 色 色综合com| 婷香五月激情视频| 色九月婷婷综合| 欧美五月婷婷| 成人中文网| 国产AV一区二区三区最新精品| 日本人妻伦在线中文字幕| 中文人妻AV久久人妻18| 国产免费性爱| 成人一区在线观看| 51成人| 一级黄色影片| 91婷婷五月天综合视频| 99热国产这里只有精品| 中文字幕综合| 夜夜爽77777妓女免费下载| 精品国产乱码久久久久久夜深人妻| 日日夜夜国产| 久久六月综合| 色婷婷操逼| 综合五月草| 国产淫熟妇| 人妻自慰高清合集| 99re在线视频| 老师的粉嫩小又紧水又多A片视频 亚洲国产精品VA在线看黑人 | 国产二区自拍| 中文字幕高清av| 国产99久| 婷婷久久丁香五月| 丁香六月视频| 俺去也五月| 天天爽天天| 天天久久狠狠色综合| 狠狠精品干练久久久无码中文字幕| 午夜精品久久久久久久99老熟妇| 五月天大香蕉视频| 久久66er久久| 99热无码| 99热免费精品| 一级内射毛片| 亚洲高清在线| 激情婷婷五月天日本系列| 婷婷五月小说| 五月丁香| 殴美激情综合网| 五月色婷婷影视在线电影| 婷婷丁香六月激情综合| 99这里热| 91青青青青青爽在线| HD久久精品视频| 91天天操天天干天天射| 激情网五月婷婷| 国产avapp 网| 国产性av| 五月色综合| 色婷婷激情五月天在线观看| 五月开心激情| 狠干综合| 2025天天爽天天摸| 综合大香蕉| 九月丁香婷婷基地| 久久精彩免费视频精彩免费视频| 天堂草在线看www| 直接看的AV| 99在线免费视频播放| 亚洲综合字幕色色| 五月亭亭网成人在线视频| 婷婷综合视频| 五月婷婷综合在线视频| 成人av在线网站| 青草激情在线| 国产真人做爰视频免费| 中文在线视频久9| 五月天天丁香婷婷在线中| 激情五婷网| www.色色五月天.com| 97人人草| 五月丁香六月色婷婷综合五月天| 久久精彩视频| www.丁香黄色五月天人与| 爱射综合| 免费观看欧美成人AA片爱我多深| 亚洲激情四谢| 青草性爱视频| 日韩欧美一级大黄网站| 大香蕉娱乐| 夜夜操少妇| 婷婷五月天av| 九九人人自拍| 91精品综合久久久久久五月丁香 | 五月丁香久久久| 全亚洲最大的婷婷五月天网站COM| 五月天婷婷在线AN| 色色欧美色色| 婷婷丁香五月天综合网| 亚洲另类在线观看| 九洲一级A片| 99爱在线精品视频免费观看| 91碰碰| 五月婷俺去也| 久久HD| 青青999| 五月婷婷激清网| 亚洲国产精品VA在线看黑人 | 色一区高清| 中文字幕激情综合| A1片久久| 成 久久| 国产真实乱了老女人视频| 91碰碰视频| 久久天天| 亚洲精品中文字幕成人片| 激情五月天色婷婷综合| 五月天激情网图片| 五月丁香综合伦理片| 五月婷婷六月激情| 丁香五月网络网络| 久久在线视频免费观看| 婷婷丁香五月天综合AV| 天天日天天摸| 日本色图综合| 综合激情在线视频| 色V狠狠的干| 狠狠99| 色九月国产| 久久综合人妻| 国产亚洲色婷婷久久99精品91 www.riverspirits.org www.hnnun.com www.changh | 成人婷婷色综合| 亚洲综合婷婷| wwwav大香蕉| 色视五月天婷婷| 欧美丰满熟妇BBB久久久| 精品无码av丁香五月激情| 激情综合网五月| 99er日韩| 亚洲色无码A片一区二区麻豆| 色婷婷综合久色AV五色最新| 丁香五月成人| 六月丁香深深爱| 欧美搡BBBBB摔BBBBB| 丁香五月第九色| 日日操日日撸| 亚洲一色色色色色色色色| 中国女人做爰A片| 都市激情蜜桃婷婷五月天| 人妻AV中文系列| 五月色欧洲| 一起草AV入口| 99热在线观看精品| 欧美性久| 激情五月小说婷婷| 中国AV性爱观看| 日韩限制级大尺度黑料泄密大尺度视频一区二区在线观看 | 激情五月综合网丁| 日日夜夜久| 久婷自拍视频| 99日本精品视频热| 99爱免费在线观看| 亚洲精品一二三| 丁香五月婷中字幕| www.99热在线观看| 国产精产国品一二三在观看| 日本在线噜噜| 日本色色网站| 99爱视频在线观看| 激情文学天天| 亚洲日韩久久婷婷伊人| 99这里有精品| 人妻狠狠操| 秋霞无码AV久久久精品小说| 亚洲国产色色| 色婷婷五月色| 亚州免费视频| 五月丁香六月激情在线| 大香蕉啪啪啪| 人妻激情久久| 99九九精品视频| 五月天天天操天天爽夜夜操| 精品一区久热| 中文字幕综合网| www.日日日.com| 91丁香婷婷综合久久欧美| 夜色综合网| 丁香婷婷五月天校园春色| 六月丁香五月婷婷| 狠狠爱婷婷丁香| 色频玖玖五月天| 人人妖人人97| 九九九热精品| 五月婷婷福利| 五月丁香婷婷啪啪| 成人国产网| 欧美综合123区| 丁香五月狠狠综合欧美| 色欲久久久久久综合网综合网| 26uuu日韩| 天天摸色吧天天摸色吧| 激情五月天久久丁香| 90色免费视频| 天天插综合网| 色5月丁香婷婷| 久久aaa| 色婷婷综合电影| 精品香蕉99久久久久网站| 六月丁香社区| 深爱 五月天| 国产黄色av| 国产亚洲精品久久久久久郑州| 九九色综合视频| 综合亚洲六月婷婷在线| 91综合色噜噜| 欧美激情一区二区三区视频| 女同激情久久av久久| 爱婷婷久久视频| 超碰在线看| 中文字幕人妻AV| 欧美性猛交AAAA片黑人 | 五月天天久久香| 丁香六月视频| 婷婷色丁香五月| 操逼视频网址| 亚洲中文字幕在线观看| 久久92| 国语精品探花| 婷婷丁香五| 亚洲成人av在线| 先锋影音男人的天堂AV| 国产AV一区二区三区日韩| 中文久久婷婷| 色婷婷五月视频| 婷丁香五月天| 欧美一级色| 激情综合网色五月| 人人操婷婷| 蜜桃麻豆WWW久久国产SEX| 九色自拍| 色婷五月天| 色欲五月婷婷| 日韩按摩二区| 26uuu国产色| www色五月| 免费看片在线观看| 色欲AVV| 蜜桃人妻无码AV天堂三区| 激情五月天视频| 丁香五月天电影| 婷婷激情六月天视频| ,99视频久久| 狠狠香蕉| 色婷婷丁香综合中文字幕| 一级七香蕉| 婷婷5月久久综合网站| 日本欧美成人片AAAA| 五月婷婷激情久久| 日本九九九九| 伊人干综合| 综合一区二区三区| 四LLL少妇BBBB槡BBBB| 婷婷五月天激情诱惑| 色噜综| 五月婷综合性中心| 六月五月久久丁香| 深爱网深爱综合网| 日本婷婷丁香五月| 天天更新天天亚洲| 天天射射夜| 激情图片婷婷| 婷婷性爱无码视频| 日日夜夜噜噜爽爽| 丁香五月六月欧美| 婷婷激情五月天在线| 五月天激情小说婷婷基地| 激情久久天天| 日日懆天天懆| 天天干狠狠艹| 激情文学五月丁香六月婷婷| 亚洲视色| 青青在线观看视频在线高清完整版| 天天做夜夜爽| 激情第四色| 婷婷五月色色| 成人视屏在线观看| 色五月丁香婷婷| 婷婷五月天福利| 强壮公让我夜夜高潮A片视频 | 激情五月色婷婷| 亚洲婷婷丁香| 午夜婷婷| WWW.桔色成人.COM| 国产黄色在线观看| 丁香婷婷久| 亚洲AV永久无码影院黑人 | 色激情五月| 五月丁香无码| 99热99成人| 日韩抽插操逼| 色五月婷婷天天操夜夜操| 狠狠色噜噜| 亚洲操B| 99热网址| 色色色综合色| 亚洲风情偷拍区| 99热超碰人| 久久婷婷东京热大香樵| 色婷婷免费观看| 这里只有精品日韩| 丁香五月天欧美| 激情99在线视频| 丁香五月偷拍| 99热思思| 91色婷婷综合久久中文字幕二区| 精品人妻伦九区久久AAA片| 91精品久久久久久综合五月天| 亚洲超碰青涩| 日本人人干| 97热精品| www.99热最新视频8| 天堂草在线观| 日本色色网站| 一本道在线电影| 国模狼狼| 色播婷婷五月天| 色。 日日日| 五月丁香六月婷婷久久| 97久久人人操| 国产毛片精品一区二区色欲黄A片| 婷婷久久婷婷色五月| 亚洲视频五区| 五月婷婷色情| 日本99在线| 一本大道熟女人妻中文字幕在线 | 99A级片| 在线观看免费视频| 综合爱久久| 另类激情五月天。| 婷婷日日夜夜| 夜夜爽天天干| 日美三级| 五月天激情综合首页| 婷婷五月图片小说视频| 色五月首页| 伊人婷婷福利网| 丁香五月91| 成人色色视频| 五月丁香啪啪综合| 九久热| 性色欲情 网站| 噜噜色噜噜网| 国产成人一区二区三区在线观看| 超碰色女人| 九九精品这里只有| 男人天堂网2017| 一区二区视频在线观看高清视频在线| 久人操| 国产一区男女| 色五月综合激情| 狠狠狠狠狠狠草| 99这里只有精品视频免费| 久久综合人妻| 国产美女69视频免费观看| 任你擦免费视频| 日日狠夜夜狠| 六月婷婷综合| 日本人人草草| 丁香五月中文字幕久色| 99久视频| 成人Av在线大片| 亚洲丁香五月在线观看| 激情婷婷六月天| 久久婷婷丁香视频网| 色播五月丁香婷婷| 99视频自拍| 月色色综合婷婷网| 五月丁香色狠狠干大屄| 人人干av| 丁香蜜臀黄色婷婷五月天| 日韩欧美一级大黄网站| 欧美片第1页 综合| 丁香五月影院| 91无码高清| 草榴视频黄色网| \\五月天婷婷激情| 色欲一区二区三区精品A片| 五月婷婷综合影院| 婷婷丁香人妻天天久久| 99无码视频| 欧美va精品va老师va| 婷婷五月色综合| 久热精彩视频98| 99热这里精| 亚洲无AV在线中文字幕| 超碰99在线观看| 99碰网站| 草榴成人影片| 成人无码精品1区2区3区免费看| 99综合免费视频| 色天天久婷婷| 色婷婷久久| 97操视频| 97碰碰在线看视频免费| 色婷婷色综合激情91| 欧美成人五月天| 在线中文AV| 婷婷六月插屄激情| 久久精品99国产精品日本| 色婷婷电影| 五月色色色| 国产脫衣舞一区二区三区| 婷婷五月色| 天综合日日夜综合7799| 色婷婷社区| 五月丁香婷婷狠狠操| 99只有这里是精品| 激情综合五月天| 婷婷操逼网| 亚洲精品中文字幕制| ...婷婷国产成人亚洲日韩| 国产亚洲成AV人片在线观黄桃| 级情九色| 色色色9| 色 免费网站视频| 九九热免费| 婷婷激情中文综合| 五月天激情国产综合婷婷婷| 99久热在线精品| 六月综合婷婷开心伊人| 久思思热视频在线观看| 少妇性按摩无码中文A片| 狠狠干五月天婷婷网| 亚洲第一综合| 成年AAAA色情| 国产亚洲在线| 五月婷婷开心丁香| 丁香五月婷婷亚洲天堂| 五月婷婷五月天| 99热在线资源| www.久久99| 亚洲五月丁香综合网| 国产SUV精品一区二区883 | 可以免费观看的AV| 婷婷激情综合色五月久久,色婷婷丁香花,丁香婷婷五月情天,久久婷婷五月综合色 | 婷婷六月视频| 丁香五月综合| 五月天婷婷午夜丁香| 国产麻豆老师在线观看| 五月丁香六月婷婷婷婷| 9在线9在线婷婷在线国产| 天天干天天干天天干| 婷婷免费无马| 久久九九国产精品怡红院| 婷婷丁香五另类网站| 久久婷婷五月综合色丁香花| 九九色热| 色色丁香五月天| 午夜69成人做爰视频| 开心五月婷婷激情网| 伊人在线大香蕉网| 97五月久久丁香婷婷| 激情五月综合色| 欧美日韩999| 玖玖热视频| 97碰| 亚洲精品久久麻豆蜜桃| 五月网在线| 91凹凸在线| 激情五月婷婷| 婷婷无码视频| 精典久久| 午夜精品人妻无码一区二区三区 | 1819岁日本MACBOOK| 99精品久久| ..真实国产乱子伦毛片| 深闺禁伦强HNP| a色色色色色| 久久久婷婷五月天| 婷婷狠狠18禁久久| 日 日干 日日做| 伊人丁香五月婷婷潮吹| 99re思思热久久| 国产精品久久久丁香五月八戒视频| 五月天色色色色色| 久久久久人妻| 99热在线这里| 亭亭五月丁香综合欧美| 色www99| 丁香六月色婷婷| 精品综合五月| 国产av基地| 五月天偷拍| 七七九色| 久久久久99精品成人片| 久久精品91视频| 在线 国产 欧美 专区| 精品色色| 干婷婷五月天| 67194中文字幕| 亚洲欧美综合在线天堂| WWW.17C.COM最新官网| 五月婷婷色男女| 91在线操| 91av传媒高清在线视频网| 精品99在线| 大香蕉院线| 九九色图| 丁J香六月首页| 久久人人妻| 丁香五月婷婷五月| 97久久香草精品视频| www,色婷婷| 丁香五月婷婷激情97| 伊人干综合| 亚洲狠狠干| 亚洲第一色色色色| 免费啪啪亚州视频| 亚州第一黄网| 亚洲字幕AV一区二区三区四区| 思思热在线视频精品| 五月婷婷六月爱| 色欲色天天香综合| 婷婷新网址| 一本大道伊人AV久久综合| 97碰人人操| 丁香婷婷十月| 久久6这里只有精品| 国产av基地| 五月婷婷 自拍| 亚洲国产中文在线视频| 大香蕉520| 99热66| 天天免费日日夜夜夜夜| www.激情五月天.com| 色婷婷激情| 丁香五月婷婷久久久| 一区二区视频在线观看高清视频在线 | 久久开心五月天激情| 午夜丁香六月婷| 综合网色| 丁香九月综合激情| 五月天婷婷色色首页| 天天草女人| 少妇做爰免费视看片| 丁香五月综合婷婷| 少妇人妻人伦A片| 人草人人| 婷婷开心激情综合五月天| 久久五月天综合视频网站| 久久综合中文| 人人干99| 99久热精品在线| 99精品久| 665566 无码| 一级AV片| 日本一级一级一级一级| 亚洲亚洲人成综合网络| 国产成人网站在线观看| 久久精品夜色噜噜亚洲a∨| 婷婷五月激情视频在线| 色噜噜婷婷| 开心色播色五月婷婷| 五月婷婷丁香五月亚洲色| 亚洲小视频免费观看| 97色色色| 97人人操人人| 99热久草| 99热亚洲| 另类激情五月| 97香蕉久久超级碰碰高清版| 97干综合网| 亚洲超级碰| 九九九九九九毛片| 激情五月份婷婷| 91精品丝袜久久久久久久久粉嫩| 欧美激情-区二区三区| 丁香五月瑟瑟| 亚洲综合色激情色五月| 婷婷激情六月中文| 中文字幕不卡网站| 丁香五月激情五月| 国产黄色大片| 激情丁香五月天图片| 婷婷激情五月天激情小说| 91人人人人人人人| 91在线观看www| 五月天久草| 91操在线观看| 亚洲无码色| 五月天激情网站| 久久九九激情五月天 | 激情五月天色网站| 色情综合网| 久久激情网| 黑人糟蹋人妻HD中文字幕| 国产在线观看不卡免费高清| 99热9| 九色综合网| 9l视频自拍九色9l黑人| 婷婷丁香五月天熟女丝袜| 亚洲一区二区无遮挡A片| 久99久热| 免费AV播放| 亚洲色色爱| 综合五月天天天天天五月| 亚洲五月天婷婷综合| 天堂草在线看www| 四月婷婷五月丁香| 99热大香蕉| 五月天婷婷综合| 亚州免费视频| 丁香五月玖玖| 免费看无码视频A级| 婷婷五月天堂| 99热20| 我要看激情五月天| 婷婷久久色| 婷色五月天| 成人国产欧美大片一区| 免费精品99| 97人人操人| 亚洲综合视频网| 色情五月丁香婷婷网| 天天摸天天高潮天天爽| 久久久免费精彩视频| 婷婷五月天久久| 极品人妻VIDEOSSS人妻| 久久这里只有精品视频15| 99热高清在线| 九九热色视频| 国精产品一区二区三区| 国产精品人成A片一区二区| 五月香婷婷| 亚洲精品一区国产欧美| 综合九九日本| 日本综合久久| 99aese| 97五月天| 亚洲妇女熟BBW| 色婷婷yy久| 人妻AV在线观看| 婷五月丁香| 久久精热| 五月婷婷六月丁香| 丁香婷婷老熟女综合网| 九月激情综合| 婷婷丁香五月精品| 五月婷视频在线| 无码人妻丰满熟妇奶水区码| 九月婷婷人人操人人舔人人爱| 五月天婷婷免费| 开心日韩丁香婷婷五月| 久久久无码A片观看免费| 天天插天天射| 欧美激情-区二区三区| 九九综合色| 婷婷五月深爱五月| 91丨九色丨大屁股| 超碰99热精品在线| 天天肏视频| 操射国产日本| 五月丁香成人网| 九月婷婷人人操人人舔人人爱| 狠狠综合久久| av五月天婷婷丁香| 99在线精品视频在线观看| 182无码| 狠狠大香婷婷爱| 五月天天天色| 熟女人妻一区二区三区免费看| 狠干综合| 久久奄也去色色网站| 色五月大| WW婷婷五月天com| 五月丁香婷婷色色| 天天日天天爱天天噪| 人妻少妇色综合| 海外网站专业操老外| 激情五月婷婷伊人| 五月丁香精品| 九九色网| 人妻系列久久久久久久久久久 | 亚洲永久免费| 无码操B| 99精品免费久久久久久久久日本 | 玖玖婷婷五月天| 99re青青草| 婷婷综合激情五月综合| 欧美在线视频99| 久久人操-久草婷婷-成人AV| 丁香六月婷婷综合网| 色婷婷伊人| 热久免费视频9| 日日噜噜久久婷婷五月天| 五月婷婷中文字幕| 久久精品日| 丁香五月激情六月综合| 夜夜干夜夜操| 九九干视频| 色五月婷婷丁香五月| AV天堂午夜精品一区二区三区| 操久久网| 丁香婷婷九月| 狠狠色婷婷综合开心影视| 日本在线免费中文com.| 99综合免费视频| 99热狠狠操| 婷婷噜噜| 在线视频婷婷| 婷婷五日b| 亚洲九九视频| 曰曰久久| 02kkkk| 碰人人97| 伊人午夜综合色啪| 99精品网站| 激情丁香久久| 色五月在线观看| enecarbon-materials.comWu染请涟系Bao护@wip1688 | 五月激情综合网| 色六月婷婷| 五月99久久| 婷婷五月激情五月丁香五月| 79色色色色| 激情AV综合| 狠狠色激情综合| 国产高清视频色拍| 九九九九九无码| 五月天激情网站| 五月婷婷xxx| 97色在线视频| 久久精品五月天| 97操碰在线视频| 久久婷中文字幕| 99精品在线观看视频| 色99xx| 久久五月天激情| 五月婷婷久久网| 久久性爱视频这里只有精品| 操操操B| 久久99这里只有精品视频| 97视频精品全国在线观看| 婷婷激情另类| 最新高清无码专区| 色婷婷视频| 国产高潮A片羞羞视频涩涩| 色爱99| 日产精品久久久久久久蜜臀| 五月婷免费视频久久久| 青柠影视免费高清电视剧| 激情五月综合视频| 免费视频WWW在线观看网站| 五月天婷婷自拍图片在线观看| 99精品国产在热久久| 婷婷激情综合无月| 国产99区| 综合色色婷婷| 丁香九月色| 日本色视| 91精品无码久久久久久五月天| 国产免费一区二区三区三州老师F1F1.CC| 丁香五月停停基地| 91人人网| 天天操天天日天天爱| 亚洲亚洲人成综合网络| 一根材五月婷成人| 五月综合久久| 丁香六月天堂| 婷婷五月天性爱视频| 99热成人精品网站| 色99在线视频| 人人草人人舔| 日本五月丁香| 亚洲182在线观看| 五月天色不卡| 9久久久久久久久久久| 婷婷丁香18| 五月婷婷丁香五月亚洲色| 丁香伍月婷电影全集| 天天婷婷| 91狠狠综合网| 婷色五月天| 婷婷丁香五月天小说| 色在线免费观看| 欧美成人精品A片免费一区99| 1024国产| 欧美人与性动交CCOO| 久久婷婷丁香六月天| 免费观看2018www黄色操逼网站| 操97免费超级视频| 青青草性爱视频| 97超碰免费超级在线观看| 狠狠草在线观看| 婷婷精品在线| 国内精品视频在线播放一区| 99热无码| 97干欧美| 91精品欧美一区二区三区| 伊人AV五月婷| 日韩在线视频网站| 国产伦亲子伦亲子视频观看| 五月婷婷丁香在线视频| 91丨九色丨老农村| 五月丁香婷婷六月| 亚洲精品五月| 99日逼视频| 蜜桃人妻无码AV天堂三区| 五月婷婷开心综合| AV在线大香蕉| 9久热| 色五月综合资源推荐| 只有精品在线观看| 久99久视频精品| 欧美久久网| 婷婷五月天视频小说| WWW久| 青青青在线视频国产| 九九99精品视频在线观看| 丁香五月婷婷激情网| 三级黄色大片视频| 2017狠狠干| 婷婷五月天伊人网在线观看视频| 九九热婷婷| 亚洲精品乱码久久久久蜜桃 | 国产99热| 激情婷婷五月天日本系列| 影音先锋噜一噜| 桃色五月婷婷| 亚洲中文字幕av| 亚洲婷婷激情五月天| 色婷五月天| 天天插操| 五月天色社区| 五月天天爽| 五月天天天操天天爽夜夜操| 99综合成人视频在线观看| 欧美三级A做爰在线观看| 六九色综合婷婷五月天| 99 频99热国里只有精品| 狠狠se| 无码一区二区三区亚洲人妻| 九月婷婷色色| 色爱终和网| 99re热精品视频国| 欧美啪啪五月天| 亚洲精品免费视频| AV操逼网| 99色免费视频| 国庆精品久久| 五月天激情四射网站| 琪琪色综合网站| 无码任你操| 亚洲va成人va成人va在线观看| 色色丁香五月婷婷| 99色网站| 五月久久| 久久伊人大香蕉| 丁香六月综合激情| 婷婷丁香五月亚洲17cao| 五月的婷婷六月丁香| 4399在线日本A片| 五月丁香A∨在线| 国产午夜精品A片一区仙踪林| 色色丁香五月天| 五月婷婷啪| 丁香久久五月天视频在线观看| 久操大香蕉| 91丁香| 日韩三级高清无码| 国产成人综合五月久久网址| 婷婷久热| www.97| 婷婷丁香色五月| www.婷婷,com| 狠狠艹狠狠艹| 婷婷色九月| 91丨九色丨老农村| 人人摸人人干| 狠狠干狠狠色| 久久国产一区二区三区| www.婷婷亚洲基地| 99网99热| 久久少妇视频| 狠狠草综合网| 伊人日日干| 五月天六月丁香| 久久激情五月| 五月婷在线影院| www.com色播五月天| 久久久久久婷| 婷婷六月激情综合| 亚洲一区免费观看| 少妇性按摩无码中文A片| 久九色| 丁香五月激情啪| 黄网在线观看免费| 五月综合激情| www.操.com|