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神經(jīng)組織消化手冊

更新時間:2016-02-16點擊次數(shù):1971

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Worthington組織消化指南

引言

組織消化酶主要用于組織培養(yǎng)和細胞生物學研究的原代細胞的分離和收集。盡管這些消化酶已經(jīng)普遍應用多年,但是對其組織消化和細胞收集的作用機制卻沒有全面的理解。  由此導致的結(jié)果便是選擇一種消化方法大多是隨機選擇或基于過去的經(jīng)驗,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)具體實驗目的修正消化方案以獲得更好的實驗結(jié)果。

細胞分離程序的目的是zui大程度的獲得功能活性的單個細胞,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、孵育時間

通常前四個原因沒辦法選擇,而剩下的原因一般由經(jīng)驗決定以獲取zui適實驗結(jié)果。

通過查閱文獻來選擇*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驗數(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適消化和細胞收集知識和方法,描述了標準的實驗步驟, 以便于制定一個*的細胞分離方案,同時列舉了一些相關(guān)的參考文獻。

下表列出多種組織的適宜消化酶(含參考文獻,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

眼

心臟

腸

腎

肝

肺

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。

 

神經(jīng)組織消化手冊

 

物種詳細來源細胞名稱消化用酶培養(yǎng)基參考文獻
鳥類小雞, 白來杭雞, 胚胎Cilary ganglion neuronsCollagenase Type 1: 0.1%PBSTemburni, M., Rosenberg, M., Pathak, N., McConnell, R. and Jacob, M.:Neuronal Nicotinic Synapse Assembly Requires the Adenomatous Polyposis Coli Tumor Suppressor Protein., J Neurosci 24, 6776, 2004
牛牛 HeartTrypsin: 0.06%25mM HEPES buffered Locke's solution, CMFTrifaro, J., Tang, R., and Novas, M.: Monolayer Co-Culture of Rat Heart Cells and Bovine Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990
(包括 大鼠)Adrenal chromaffin 
 Paraneurons
牛牛Microvascular endothelialCollagenase/Dispase: 0.1%MEMBowman, P., Betz, A., and G大stein, G.: Primary Culture of Microvascular Endothelial Cells From Bovine Retina, In Vitro 18 (7), 626, 1982
牛牛犢OligodendrogliaTrypsin: 0.1%(see reference)Poduslo, S., Miller, K., and McKhann, G.: Metabolic Properties of Maintained Oligodendroglia Purified from Brain, J Biol Chem 253, 1592, 1978
(包括 小羊羔)Neural
雞雞, 白來杭雞, 胚胎, 17-21 天NeuronsPapain: 40 u/mlHEPESRaman, I., and Trussell, L.: The Kinetics of the Response to Glutamate and Kainte in Neurons of the Avian and Cochlear Nucleus, Neuron 9, 173, 1992
雞小雞, 胚胎, 10-14 天Flat, retinaTrypsin: 0.1%Tyrode's solution, CMFMoyer, M., Bullrich, F., and Sheffield, J.: Emergence of Flat Cells From Glia in Stationary Cultures of 胚胎Chick Neural Retina, In Vitro Cell Dev Biol26, 1073, 1990
雞小雞, 白來杭雞, 胚胎Cerebral neuronsTrypsin: Trypsin: 0.125-0.25%DMEMCoates, P., and Nathan, R.: Feasibility of Electrical Recordings From Unconnected Vertebrate CNS Neurons Cultured in a Three-Dimensional Extracellular Matrix, J Neurosci Methods 20, 203, 1987
雞小雞, 白來杭雞, 胚胎Spinal cordTrypsin: 0.05%Phosphate bufferSchnaar, R., and Schaffner, A.: Separation of cell types from 胚胎chicken and rat spinal cord: characterization of motoneuron-enriched fractions, J Neurosci 1, 204, 1981
(包括SD大鼠, 胚胎)(see reference)
雞小雞Dorsal root ganglion neuronsTrypsin: 0.1%Puck's saline, CMFChoi, D., and Fiscbach, G.: GABA Conductance of Chick Spinal Cord and Dorsal Root Ganglion Neurons in Cell Culture, J Neurophysiol 45, 605, 1981
Spinal cord
雞小雞, 白來杭雞 胚胎, 8 天Neurons, gangliaTrypsin: 0.25%HBSS, CMFBottenstein, J., Skaper, S., Varon, S., and Sato, G.: Selective Survival of Neurons from Chick 胚胎 Sensory Ganglionic Dissociates Utilizing Serum-Free Supplemented Medium, Exp Cell Res 125, 183, 1980
雞雞, 白來杭雞, 種蛋Ciliary ganglion neuronsTrypsin: 0.25%Eagle's MEMTuttle, J., Suszkiw, J., and Ard, M.: Long-Term Survival and Development of Dissociated Parasympathetic Neurons in Culture, Brain Res 183, 161, 1980
雞雞, 胚胎, 9-10 天大Dorsal root ganglia neuronsCollagenase: 0.01%Eagle's MEMMudge, A., Leeman, S., and Fischbach, G.: Enkephalin Inhibits Release of Substance P From Sensory Neurons in Culture and Decreases Action Potential Duration, Proc Natl Acad Sci U S A 76 (1), 526, 1979
雞小雞胚胎Ganglion chains, sympathetic gangliaTrypsin: 0.25%Krebs PhosphosalineMcCarthy, K., and Partlow, L.: Preparation of Pure Neuronal and Non-Neuronal Cultures From 胚胎Chick Sympathetic Ganglia: A New Method Based on Both Differential Cell Adhesiveness and the Formation of Homotypic Neuronal Aggregates, Brain Res 114, 391, 1976
雞小雞, 白來杭雞, 胚胎, 7 天Neural retinaCollagenase: 0.25%HBSSWiseman, L., and Hammond, W.: The Reacquisition of Cell Adhesiveness Following Tissue Disaggregation by Eleven Different Agents, J Exp Zool 197, 429, 1976
Elastase: 0.2%
Hyaluronidase: 1.0%
Papain: 1.0%
Protease: 0.1%
Trypsin: 0.05%
魚魚, 斑馬魚Motor neuronsCollagenase Type 2: 0.1%See referenceSakowski, S., Lunn, J., Busta, A., Palmer, M., Dowling, J. and Feldman, E.: A Novel Approach to Study Motor Neurons from Zebrafish 胚胎s and Larvae in Culture., J Neurosci Methods 205, 277-82, 2012
魚 Dania rerio斑馬魚,  胚胎Rohon-Beard neuronsTrypsin: 0.2%L-15/HepesWon, Y., Ono, F. and Ikeda, S.: Identification and Modulation of Voltage-Gated Ca2+ Currents in Zebrafish Rohon-Beard Neurons., J Neurophysiol 105, 442-53, 2011
魚Dania rerio斑馬魚, 胚胎NeuronsPDS kit: with modificationsEBSSCerda, G., Hargrave, M. and Lewis, K.: RNA Profiling of FAC-sorted Neurons from the Developing Zebrafish Spinal Cord., Dev Dyn 238, 150-61, 2009
魚黑色的魅影kni克e魚, 成年的(黑魔鬼魚)Neurons, spinal cordTrypsin: 0.4%PBS, CMFAnderson, M.J.: Differences in Growth of Neurons from Normal and Regenerated eost Spinal Cord in vitro, In Vitro Cell Dev Biol 29A, 145, 1993
青蛙非洲爪蟾蜍, 胚胎NeuronCollagenase Type 1: 0.1%Steinberg's solutionTakahashi, T., Nakajima, Y., Hirosawa, K., Nakajima, S., and Onodera, K.: Structure and Physiology of Developing Neuromuscular Synapses in Culture, J Neurosci 7, 473, 1987
豚鼠豚鼠, 初生Neuron, entericTrypsin: 0.125%Medium 199Jessen, K, McConnell, J., Purves, R., Burnstock, G., and Chamley-Campbell, J.: Tissue Culture of Mammalian Enteric Neurons, Brain Res 152, 573, 1978
馬馬, 雄性Vomeronasal organ neuronsCollagenase Type 1: 0.02%PBSLiman ER: Regulation by voltage and adenine nucleotides of a Ca2+-activated cation channel from hamster vomeronasal sensory neurons, J Physiol 548, 777, 2003
Trypsin: 0.02%
人類人類Neural, variousPapain: 12 u/mlsee referencePanchision David M, Chen Hui-Ling, Pistollato Francesca, Papini Daniela, Ni Hsiao-Tzu, Hawley Teresa S: Optimized flow cytometric analysis of central nervous system tissue reveals novel functional relationships among cells expressing CD133, CD15, and CD24, Stem Cells 25, 1560-70, 2007
Trypsin: see reference
Collagenase/Dispase: see reference
人類人類, 9 個月NeuronsCollagenase Type 4: 1.33%DMEM/F12Dietrich J, Lacagnina M, Gass D, Richfield E, Mayer-Proschel M, Noble M, Torres C, Proschel C.: EIF2B5 mutations compromise GFAP+ astrocyte generation in vanishing white matter leukodystrophy, Nat Med 11, 277, 2005
Papain: 0.07 u/ml
Neutral Protease: 1 mg/ml
人類人類, 26 周Neural progenitor cellsPapain: 2.5 u/mlDMEM/F-12Fuja TJ, Schwartz PH, Darcy D, Bryant PJ.: Asymmetric localization of LGN but not AGS3, two homologs of Drosophila pins, in dividing human neural progenitor cells, J Neurosci Res 75, 782, 2004
Deoxyribonuclease I: 250 u/ml
Neutral Protease: 1 u/ml
人類人類, 成年人Ventricular epithelialPapain: 11.4 u/mlDMEM/F12Roy NS, Benraiss A, Wang S, Fraser RA, Goodman R, Couldwell WT, Nedergaard M, Kawaguchi A, Okano H, G大man SA: Promoter-targeted selection and isolation of neural progenitor cells from the adult human ventricular zone, J Neurosci Res 59, 321, 2000
Deoxyribonuclease I: 10 u/ml
人類人類, 5-65 歲Retinal pigment epithelial (RPE)Trypsin: 0.25%HBSSVon Recum, H., Okano, T., Kim, S, and Bernstein, P.: Maintenance of Retinoid Metabolism in Human Retinal Pigment Epithelium Cell Culture, Exp Eye Res 69, 97, 1999
人類人類Dorsal root ganglion neuronsCollagenase Type 1: 0.2%DMEM/F-12Dib-Hajj SD, Tyrrell L, Cummins TR, Black JA, Wood PM, Waxman SG.:Two tetrodotoxin-resistant sodium channels in human dorsal root ganglion neurons, F.E.B.S. Lett. 462, 117, 1999
Neutral Protease: 0.5%
昆蟲果蠅, 幼蟲NeuroblastsCollagenase Type 1: 0.1%Rinaldini solutionBerger, C., Harzer, H., Burkard, T., Steinmann, J., van der Horst, S., Laurenson, A., Novatchkova, M., Reichert, H. and Knoblich, J.: FACS Purification and Transcriptome Analysis of Drosophila Neural Stem Cells Reveals a Role for Klumpfuss in Self-Renewal., Cell Rep 2, 407, 2012
Papain: 0.1%
昆蟲果蠅DendritesCollagenase: 0.05%HBSSSanchez-Soriano, N., Bottenberg, W., Fiala, A., Haessler, U., Kerassoviti, A., Knust, E., Lohr, R. and Prokop, A.: Are Dendrites in Drosophila Homologous to Vertebrate Dendrites?, Dev Biol 288, 126, 2005
Neutral Protease: 0.2%
昆蟲雙斑蟋Giant interneuronsCollagenase: 0.05%Leibovitz's L15Kloppenburg, P. and Horner, M.: Voltage-Activated Currents in Identified Giant Interneurons Isolated from Adult Crickets Gryllus Bimaculatus, J Exp Biol201 (Pt 17), 2529, 1998
Neutral Protease: 0.2%
小鼠小鼠DRG neuronsCollagenase Type 2: 0.1%DMEM/F-12Lee, B., Cho, H., Jung, J., Yang, Y., Yang, D. and Oh, U.: Anoctamin 1 Contributes to Inflammatory and Nerve-Injury Induced Hypersensitivity., Mol Pain 10, 5, 2014
Trypsin: 0.25%
小鼠小鼠, 新生的Schwann cellsCollagenase: 0.05-0.1%DMEMStettner, M., Lohmann, B., Wolffram, K., Weinberger, J., Dehmel, T., Hartung, H., Mausberg, A. and Kieseier, B.: Interleukin-17 Impedes Schwann Cell-Mediated Myelination., J Neuroinflammation 11, 63, 2014
Trypsin: 0.125-0.25%
小鼠小鼠OligodendrocytesPapain: 0.1%HBSSHarris, M., Hulseberg, P., Ling, C., Karman, J., Clarkson, B., Harding, J., Zhang, M., Sandor, A., Christensen, K., Nagy, A., Sandor, Matyas,S. and Fabry, Z.: Immune Privilege of the CNS is not the Consequence of Limited Antigen Sampling., Sci Rep 4, 4422, 2014
Deoxyribonuclease I: 0.0002%
小鼠小鼠hippocampal neuronsPDS kit: per instructionsDMEMWilliams, E., Zhong, X., Mohamed, A., Li, R., Liu, Y., Dong, Q., Ananiev, G., Mok, J., Lin, B., Lu, J., Chiao, C., Cherney, R., Li, H., Zhang, S. and Chang, Q: Mutant Astrocytes Differentiated from Rett Syndrome Patients-Specific iPSCs Have Adverse Effects on Wild-Type Neurons., Hum Mol Genet23, 2968-80, 2014
小鼠小鼠NeurosperesPapain: 0.1%NeuroCult NSCStuart, M., Corrigan, F. and Baune, B.: Knockout of CXCR5 Increases the Population of Immature Neural Cells and Decreases Proliferation in the Hippocampal Dentate Gyrus., J Neuroinflammation 11, 31, 2014
Deoxyribonuclease I: 0.1%
小鼠小鼠, 4-6 周SchwannCollagenase Type 1: 0.01%DMEMMasaki, T., Qu, J., Cholewa-Waclaw, J., Burr, K., Raaum, R. and Rambukkana, A.: Reprogramming Adult Schwann Cells to Stem Cell-Like Cells by Leprosy Bacilli Promotes Dissemination of Infection., Cell 152, 51, 2013
Trypsin: 0.125%
小鼠小鼠Oligodendrocyte precursor cellsPapain: 0.09%DMEMMedina-Rodriguez, E., Arenzana, F., Bribian, A. and de Castro, F.:Protocol to Isolate a Large Amount of Functional Oligodendrocyte Precursor Cells from the Cerebral Cortex of Adult Mice and Humans., PLoS ONE 8, e81620, 2013
小鼠小鼠, 6-8 周Neural precursorsTrypsin: 0.13%see referenceBabona-Pilipos, R., Popovic, M. and Morshead, C.: A Galvanotaxis Assay for Analysis of Neural Precursor Cell Migration Kinetics in an Externally Applied Direct Current Electric Field., J Vis Exp 68, e4193, 2012
Hyaluronidase: 0.08%
小鼠小鼠, 成年的Retinal neuronsPapain: 10-20 ulHBSSGoetz, J. and Trimarchi, J.: Single-Cell Profiling of Developing and Mature Retinal Neurons., J Vis Exp 62, e3824, 2012
小鼠小鼠NeuronsPapain: 100 ulDMEMLi, Y., Roy, B., Wang, W., Zhang, L., Sampson, S. and Lin, D.: Imaging pHluorin-Tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons., J Vis Exp 69, e4450, 2012
小鼠小鼠, P4-5NeuroshereTrypsin: 0.25%Pro-NZiegler, A., Schneider, J., Qin, M., Tyler, W., Pintar, J., Fraidenraich, D., Wood, T and Levison, S.: IGF-II Promotes Stemness of Neural Restricted Precursors., Stem Cells 30, 1265, 2012
Papain: 100 u
Deoxyribonuclease I: 0.025%
小鼠小鼠, 6-8 周Neural stem cellsPapain: 0.01%DMEM/F12Bracko, O., Singer, T., Aigner, S., Knobloch, M., Winner, B., Ray, J., Clemenson, G., Suh, H., Couillard-Despres, S., Aigner, L., Gage, F. and Jessberger, S.: Gene Expression Profiling of Neural Stem Cells and their Neuronal Progeny Reveals IGF2 as a Regulator of Adult Hippocampal Neurogenesis., J Neurosci 32, 3376-87, 2012
Neutral Protease: 0.1%
Deoxyribonuclease I: 0.01%
小鼠小鼠, 胚胎, 14 天Spinal cord neuronsPapain: 0.05%PBS/DMEMPollari, E., Savchenko, E., Jaronen, M., Kanninen, K., Malm, T., Wojciechowski, S., Ahtoniemi, T., G大steins, G., Giniatullina, R., Giniatullin, R., Koistinaho, J. and Magga, J.: Granulocyte Colony Stimulating Factor Attenuates Inflammation in a Mouse Model of Amyotrophic Lateral Sclerosis., J Neuroinflammation 8, 74, 2011
Deoxyribonuclease I: 0.004%
小鼠小鼠, 1-2 天Oligodendrocytes, dorsal root gangliaPapain: 0.15%DMEMO'Meara, R., Ryan, S., Colognato, H. and Kothary, R.: Derivation of Enriched Oligodendrocyte Cultures and Oligodendrocyte/Neuron Myelinating Co-Cultures from Post-Natal Murine Tissues., J Vis Exp 54, 3324, 2011
Deoxyribonuclease I: 0.006%
小鼠小鼠, 胚胎MotorneuronsTrypsin: 0.025%HBSSConrad, R., Jablonka, S., Sczepan, T., Sendtner, M., Wiese, S. and Klausmeyer, A.: Lectin-Based Isolation and Culture of Mouse 胚胎Motoneurons., J Vis Exp 55, e3200, 2011
小鼠小鼠, 4-8 周Olfactory ensheathingCollagenase: 0.15%DMEMRadtke, C., Sasaki, M., Lankford, K., Gallo, V. and Kocsis, J.: CNPase Expression in Olfactory Ensheathing Cells., J Biomed Biotechnol 2011, 608496, 2011
Papain: 12 u/ml
小鼠小鼠Dentate Gyrus Neural PrecursorPapain: 2.5 u/mlNeurobasal ABabu, H., Claasen, J., Kannan, S., Runker, A., Palmer, T. and Kempermann, G.: A Protocol for Isolation and Enriched Monolayer C*tion of Neural Precursor Cells from Mouse Dentate Gyrus.,Front Neurosci Vol. 5,, , 89, 2011
Neutral Protease: 1.0 u/ml
Deoxyribonuclease I: 250 u/ml
小鼠小鼠Hippocampal and retinal neuronsPapain: 1%HBSSBrown, J., Gianino, S. and Gutmann, D.: Defective cAMP Generation Underlies the Sensitivity of CNS Neurons to Neurofibromatosis-1 Heterozygosity, J Neurosci 30, 5579, 2010
Deoxyribonuclease I: 5 u/ml
小鼠小鼠CNS leukocytesCollagenase Type 4: 300 u/mlHBSSSayed, B., Christy, A., Walker, M. and Brown, M.: Meningeal Mast Cells Affect Early T Cell Central Nervous System Infiltration and Blood-Brain Barrier Integrity Through TNF: a Role for Neutrophil Recruitment?, J Immunol 184, 6891-900, 2010
小鼠小鼠, 成年的Spinal microgangliaPapain: 0.2%Hibernate AYip, P., Kaan, T., Fenesan, D. and Malcangio, M.: Rapid Isolation and Culture of Primary Microglia from Adult Mouse Spinal Cord., J Neurosci Methods 183, 223-37, 2009
小鼠小鼠, 胚胎Dopaminergic neuronsTrypsin: 0.1%DMEMRadad Khaled, Gille Gabriele, Rausch Wolf-Dieter: Dopaminergic neurons are preferentially sensitive to long-term rotenone toxicity in primary cell culture, Toxicol In Vitro 22, 68-74, 2008
Deoxyribonuclease I: 0.02%
小鼠小鼠, 新生的DRG neuronsCollagenase: 0.2%Ham's F12Pedrola, L., Espert, A., Valdes-Sanchez, T., Sanchez-Piris, M., Sirkowski, E., Scherer, S., Farinas, I., and Palau, F.: Cell Expression of GDAP1 in the Nervous System and Pathogenesis of Charcot-Marie-Tooth Type 4A Disease.,J Cell Mol Med 12, 679, 2008
Trypsin: 0.05%
小鼠小鼠, 6 個月Neurons, neurospheresPapain: 0.2%HibernateBrewer Gregory J, Torricelli John R: Isolation and culture of adult neurons and neurospheres, Nat Protoc 2, 1490-8, 2007
小鼠小鼠Neural, variousPapain: 12 u/mlsee referencePanchision David M, Chen Hui-Ling, Pistollato Francesca, Papini Daniela, Ni Hsiao-Tzu, Hawley Teresa S: Optimized flow cytometric analysis of central nervous system tissue reveals novel functional relationships among cells expressing CD133, CD15, and CD24, Stem Cells 25, 1560-70, 2007
Trypsin: see reference
Collagenase/Dispase: see reference
小鼠小鼠, 6-8 周Sensory neurons, DRGPapain: 20 u/mlHBSSMalin, S., Davis, B. and Molliver, D.: Production of Dissociated Sensory Neuron Cultures and Considerations for their use in Studying Neuronal Function and Plasticity, Nat Protoc 2, 152, 2007
Collagenase Type 2: 0.4%
Neutral Protease: 0.46%
小鼠小鼠, C57BL/6, 1 歲NeuronsPapain: 0.2%DMEMEide, L, and McMurray, C: Culture of Adult Mouse Neurons, Biotechniques38(1), 99-104, 2005
小鼠小鼠, 成年的Brain and spinal cord cellsTrypsin: 0.25%PBSGonzalez John M, Bergmann Cornelia C, Fuss Babette, Hinton David R, Kangas Cindy, Macklin Wendy B, Stohlman Stephen A: Expression of a dominant negative IFN-gammareceptor on mouse oligodendrocytes, Glia 51, 22-34, 2005
小鼠小鼠, 新生的NeuronsPDS kit: per instructionsEBSSGill JC, Moenter SM, Tsai PS: Developmental regulation of gonadotropin-releasing hormone neurons by fibroblast growth factor signaling,Endocrinology 145, 3830, 2004
小鼠小鼠, C57BLCerebellar granule cell precursorsPapain: 16.5 u/mlDulbecco's PBSOkano-Uchida T, Himi T, Komiya Y, and Ishizaki Y: Cerebellar granule cell precursors can differentiate into astroglial cells, Proc Natl Acad Sci U S A 101, 1211, 2004
Deoxyribonuclease I: 0.008%
小鼠小鼠, 1-3 天Neurons, gangliaPapain: 20 u/mlHBSSSavchenko V, Sung U, Blakely RD.: Cell surface trafficking of the antidepressant-sensitive norepinephrine transporter revealed with an ectodomain antibody, Mol Cell Neurosci 24, 1131, 2003
Deoxyribonuclease I: 100 u/ml
Collagenase: 0.3%
Trypsin: 0.05%
小鼠小鼠,雄性Trigeminal senesory neuronsPapain: 20 u/mlHEPES buffered salineRoberts, L., MacDonald, C.and Mark, C.: Anandamide is a Partial Agonist at Native Vanilloid Receptors in Acuy Isolated Mouse Trigeminal Sensory Neurons., Br J Pharmacol 137, 421, 2002
小鼠小鼠, 出生后一天的NeuronsTrypsin: 0.25%NGF-containing mediumDeshmukh, M., Kuida, K., and Johnson Jr., E.: Caspase Inhibition Extends the Commitment to Neuronal Death Beyond Cytochrome c Release to the Point of Mitochondrial Depolarization, J Cell Biol 150 (1), 131, 2000
小鼠小鼠 胚胎Neurons, DRG and SCGTrypsin: 0.25%L-15 mediumLee, K., Davies, A., and Jaenisch, R.: P75-Eficient 胚胎Dorsal Root Sensory and Neonatal Sympathetic Neurons Display a Decreased Sensitivity to NGF, Development 120, 1027, 1994
小鼠小鼠, 65 天Neurons, neuronal precursorsTrypsin: 0.1%DMEMRichards L., Kilpatrick T., and Bartlett P.: De Novo Generation of Neuronal Cells from the Adult Mouse Brain, Proc Natl Acad Sci U S A 89, 8591, 1992
Deoxyribonuclease I: 0.001%
小鼠小鼠, CD-1, neonateNeurons, dorsal root ganglionTrypsin: 0.25%HBSSQuinn, S. and De Boni, U.: Enhanced Neuronal Regeneration by Retinoic Acid of Murine Dorsal Root Ganglia and of Fetal Murine and human Spinal Cord in vitro, In Vitro Cell Dev Biol 27, 55, 1991
小鼠小鼠, 胎兒PrecursorTrypsin: 0.5%PBSKitani, H., Shiurba, R., Sakakura, T., Tomooka, Y.: Isolation and Characterization Of Mouse Neural Precursor Cells in Primary Culture, In Vitro Cell Dev Biol 27, 615, 1991
小鼠小鼠 (SWR or CF1), 1-3 月sPapillae, taste receptorPronase E: 0.15%Carbonate-Phosphate bufferSpielman, A., Mody, I., Brand, J., Whitney, G., MacDonald, J., and Salter, M.: A Method for Isolating and Patch-Clamping Single Mammalian Taste Receptor Cells, Brain Res 503, 326, 1989
(see reference)
小鼠小鼠, 新生的PNS test neuronsTrypsin: 0.08%Eagle's Basal Medium (see reference)Varon, S., Skaper, S., Barbin, G., Selak, I., and Manthorpe, M.: Low Molecular Weight Agents Support Survival of Cultured Neurons From the Central Nervous System, J Neurosci 4 (3), 654, 1984
(包括 小雞)
小鼠小鼠 (白變種實驗室老鼠), 成年的Neurons, spinal cordCollagenase Type 3: 0.25%Hank's BSS, CMFEagleson, K. and Bennett, M.: Survival of Purified Motor Neurons In Vitro: Effects of Skeletal Muscle-Conditioned Medium, Neurosci Lett 38, 187, 1983
小鼠小鼠, 0-30 天NeuralTrypsin NF 1:250: 50 0.25%BSSShrier, B., Wilson, S., and Nirenberg, M.: Cultured Cell Systems and Methods for Neurobiology, Vol. 32,, , 765, 1974
綿羊羊羔OligodendrogliaTrypsin: 0.1%(see reference)Poduslo, S., Miller, K., and McKhann, G.: Metabolic Properties of Maintained Oligodendroglia Purified from Brain, J Biol Chem 253, 1592, 1978
(包括 小牛)Neural
豬豬, 成年的, 60-100 公斤Superior cervical gangliaPapain: 2 u/mlHBSSSi ML, Lee TJ.: Presynaptic alpha7-nicotinic acetylcholine receptors mediate nicotine-induced nitric oxidergic neurogenic vasodilation in porcine basilar arteries, J Pharmacol Exp Ther 298, 122, 2001
Collagenase: 0.12%
Neutral Protease: 0.48%
鵪鶉鵪鶉Neural crestTrypsin: 0.05%MEM, HBSSSieber-Blum, M., and Cohen, A.: Clonal Analysis of Quail Neural Crest Cells: They are Pluripotent and Differentiate in Vitro in the Absence of Noncrest Cells, Dev Biol 80, 96, 1980
大鼠大鼠, 胚胎和 新生的Cortical neurons, DRGPDS kit: per instructionsNeurobasalRheaume, C., Cai, B., Wang, J., Fernandez-Salas, E., Aoki, K., Francis, J. and Broide, R.: A Highly Specific Monoclonal Antibody for Botulinum Neurotoxin Type A-Cleaved SNAP25., Toxins 7, 2354-70, 2015
Collagenase Type 1: 0.17%
大鼠大鼠, 胚胎Cerebral neuronsPapain: 2 u/mlDMEM-PBSOyanagi, K., Tashiro, T. and Negishi, T.: Cell-Type-Specific and Differentiation-Status-Dependent Variations in Cytotoxicity of Tributyltin in Cultured Rat Cerebral Neurons and Astrocytes., J Toxicol Sci 40, 459-68, 2015
Deoxyribonuclease I: 0.01%
大鼠SD大鼠, 6-9 周Dorsal Root GangliaCollagenase Type 3: 0.2%HBSSMurayama, C., Watanabe, S., Nakamura, M. and, Norimoto, H.: Inhibitory Activity of Yokukansankachimpihange against Nerve Growth Factor-Induced Neurite Growth in Cultured Rat Dorsal Root Ganglion Neurons.,Molecules 20, 14959-69, 2015
大鼠大鼠Dorsal root ganglion neuronsCollagenase Type 2: 0.15%HBSSYu, R., Seymour, V., Berecki, G., Jia, X., Akca, M., Adams, D., Kaas, Q. and Craik, D.: Less is More: Design of a Highly Stable Disulfide-Deleted Mutant of Analgesic Cyclic 偽-Conotoxin Vc1.1., Sci Rep 5, 13264, 2015
大鼠大鼠, 新生的Schwann cellsCollagenase: 0.05-0.1%DMEMStettner, M., Lohmann, B., Wolffram, K., Weinberger, J., Dehmel, T., Hartung, H., Mausberg, A. and Kieseier, B.: Interleukin-17 Impedes Schwann Cell-Mediated Myelination., J Neuroinflammation 11, 63, 2014
Trypsin: 0.125-0.25%
大鼠大鼠, 1-3 天Hippocampal NeuronsPapain: 25 u/mlL-15Thurner, P., Gsandtner, I., Kudlacek, O., Choquet, D., Nanoff, C., Freissmuth, M. and Zezula, J.: A Two-State Model for the Diffusion of the A2A Adenosine Receptor in Hippocampal Neurons: Agonist-Induced Switch to Slow Mobility is Modified by Synapse-Associated Protein 102 (SAP102)., J Biol Chem 289, 9263-74, 2014
大鼠SD大鼠, 胚胎Hypothalamic neuronalPDS kit: per instructionsNeurobasal ALoktev, A. and Jackson, P.: Neuropeptide Y Family Receptors Traffic via the Bardet-Biedl Syndrome Pathway to Signal in Neuronal Primary Cilia., Cell Rep 5, 1316-29, 2013
大鼠SD大鼠, 175-200 克Trigeminal gangliaPapain: 20 u/mlHBSSYan, J., Melemedjian, O., Price, T. and Dussor, G.: Sensitization of Dural Afferents Underlies Migraine-Related Behavior Following Meningeal Application of Interleukin-6 (IL-6)., Mol Pain 8, 6, 2012
Collagenase Type 2: 0.3%
大鼠大鼠, E19-21 and P1-3Sympathetic neuronsCollagenase Type 2: 0.1%DMEM/F-12Ghogha, A., Bruun, D. and Lein, P.: Inducing Dendritic Growth in Cultured Sympathetic Neurons., J Vis Exp 61, e3546, 2012
Neutral Protease: 0.5%
大鼠大鼠, 20-30 天Dorsal Root GanglionPapain: 20 u/mlL-15Bosmans, F., Puopolo, M., Martin-Eauclaire, M., Bean, B., Swartz, K.:Functional Properties and Toxin Pharmacology of a Dorsal Root Ganglion Sodium Channel Viewed through its Voltage Sensors., J Gen Physiol 138, 59, 2011
Collagenase Type 1: 0.3%
Neutral Protease: 0.4%
大鼠SD大鼠, 200-300 克Dorsal root ganglia, fibroblastCollagenase: 0.125%DMEMEast, E., de Oliveira, D., G大ing, J. and Phillips, J.: Alignment of Astrocytes Increases Neuronal Growth in Three-Dimensional Collagen Gels and is Maintained Following Plastic Compression to Form a Spinal Cord Repair Conduit., Tissue Eng Part A 16, 3173, 2010
大鼠大鼠,食魚貂, 7-21 月Hippocampal neuronsPapain: 0.2%Hibernate AChen, N., Newcomb, J., Garbuzova-Davis, S., Davis Sanberg, C., Sanberg, P. and Willing, A.: Human Umbilical Cord Blood Cells Have Trophic Effects on Young and Aging Hippocampal Neurons in Vitro., Aging Dis 1, 173, 2010
大鼠SD大鼠, E18NeuronalPapain: 0.2%Neurobasal EPeltier, D., Simms, A., Farmer, J and Miller, D.: Human Neuronal Cells Possess Functional Cytoplasmic and TLR-Mediated Innate Immune Pathways Influenced by Phosphatidylinositol-3-Kinase Signaling, J Immunol 184, 7010, 2010
大鼠SD大鼠, 7 天Cerebellar granule neuronsPDS kit: per instructionsPBSTanaka, S., Shaikh, I., Chiocca, E. and Saeki, Y.: The Gs-Linked Receptor GPR3 Inhibits the Proliferation of Cerebellar Granule Cells During Postnatal Development., PLoS ONE 4, e5922, 2009
大鼠SD大鼠, 胚胎天 18Hippocampal neuronsPapain: 20 u/mlNeurobasal/B27Liu, Y., Yohrling, G., Wang, Y., Hutchinson, T., Brenneman, D., Flores, C., Zhao, B.: Carisbamate, a Novel Neuromodulator, Inhibits Voltage-Gated Sodium Channels and Action Potential Firing of Rat Hippocampal Neurons.,Epilepsy Res 83, 66, 2009
大鼠大鼠, 胚胎Hypothalamic neurons and gliasPapain: 0.5 u/mlDMEMYokosuka Makoto, Ohtani-Kaneko Ritsuko, Yamashita Kayoko, Muraoka Daisuke, Kuroda Yoichiro, Watanabe Chiho: Estrogen and environmental estrogenic chemicals exert developmental effects on rat hypothalamic neurons and glias, Toxicol In Vitro 22, 1-9, 2008
大鼠 Wistar大鼠, 7 天Superior cervical ganglionCollagenase: 0.05%L-15Sakisaka, T., Yamamoto, Y., Mochida, S., Nakamura, M., Nishikawa, K., Ishizaki, H., Okamoto-Tanaka, M., Miyoshi, J., Fujiyoshi, Y., Manabe, T. and Takai, Y.: Dual Inhibition of SNARE Complex Formation by Tomosyn Ensures Controlled Neurotransmitter Release., J Cell Biol 183, 323, 2008
大鼠大鼠, 新生的Dopamine neuronsPapain: 20 u/mlPBSFrank, L., Caldera-Siu, A. and Pothos, E.: Primary Dissociated Midbrain Dopamine Cell Cultures from Rodent Neonates., J Vis Exp 21, 820, 2008
大鼠 Wistar大鼠, 4 天Hippocampal neuronsPDS kit: per instructionsNeurobasal AObradovic Darja, Gronemeyer Hinrich, Lutz Beat, Rein Theo: Cross-talk of vitamin D and glucocorticoids in hippocampal cells, J Neurochem 96, 500-9, 2006
大鼠SD大鼠, 新生的astrocytesPDS kit: per instructionsDMEMLacroix-Fralish, M., Tawfik, V., Nutile-McMenemy, N., Harris, B. and Deleo, J.: Differential Regulation of Neuregulin 1 Expression by Progesterone in Astrocytes and Neurons., Neuron Glia Biol 2, 227, 2006
大鼠大鼠, 2 月Spinal cord progenitor cells PDS kit: see referenceNeurobasal AMothe, A., Kulbatski, I., Van Bendegem, R., Lee, L., Kobayashi, E., Keating, A., and Tator, C.: Analysis of Green Fluorescent Protein Expression in Transgenic Rats for Tracking Transplanted Neural Stem/Progenitor Cells, J Histochem Cytochem 53(10), 1215, 2005
Hybrid toxins
大鼠SD大鼠, E19Dorsal root gangliaPDS kit: per instructionsMEM/Ham's F12Gavva NR, Tamir R, Qu Y, Klionsky L, Zhang TJ, Immke D, Wang J, Zhu D, Vanderah TW, Porreca F, Doherty EM, Norman MH, Wild KD, Bannon AW, Louis JC, Treanor JJ.: AMG 9810 [(E)-3-(4-t-butylphenyl)-N-(2,3-dihydrobenzo[b][1,4] dioxin-6-yl)acrylamide], a novel vanilloid receptor 1 (TRPV1) antagonist with antihyperalgesic properties, J Pharmacol Exp Ther 313, 474, 2005
大鼠 Wistar大鼠, E15Vomeronasal receptor neuronsCollagenase/Dispase: 0.1%DMEM/F12Moriya-Ito K, Osada T, Ishimatsu Y, Muramoto K, Kobayashi T, Ichikawa M: Maturation of vomeronasal receptor neurons in vitro by coculture with accessory olfactory bulb neurons, Chem Senses 30, 111, 2005
Papain: 0.5 u/ml
大鼠SD大鼠, 1-2 天Cortical astrocytesPapain: see referenceDMEMFloyd Candace L, Gorin Fredric A, Lyeth Bruce G: Mechanical strain injury increases intracellular sodium and reverses Na+/Ca2+ exchange in cortical astrocytes, Glia 51, 35-46, 2005
大鼠SD大鼠, 0-2 天Superior cervical ganglionCollagenase Type 4: 20 u/mlDMEMPedraza Carlos E, Podlesniy Petar, Vidal Noemi?, Arevalo Juan Carlos, Lee Ramee, Hempstead Barbara, Ferrer Isidre, Iglesias Montse, Espinet Carme: Pro-NGF isolated from the human brain affected by Alzheimer's disease induces neuronal apoptosis mediated by p75NTR, Am J Pathol 166, 533-43, 2005
Trypsin: 0.25%
大鼠SD大鼠,雄性, 5-8 周Trigeminal neuronsPapain: 20 u/mlCMF HanksConnor, M, Naves, L. and McCleskey, E.: Contrasting Phenotypes of Putative Proprioceptive and Nociceptive Trigeminal Neurons Innervating Jaw Muscle in Rat., Mol Pain 1, 31, 2005
Collagenase: 0.3%
大鼠大鼠, 胎兒Brainstem and cortical neuronsPDS kit: per instructionsDMEMLovshin JA, Huang Q, Seaberg R, Brubaker PL, Drucker DJ:Extrahypothalamic expression of the glucagon-like peptide-2 receptor is coupled to reduction of glutamate-induced cell death in cultured hippocampal cells, Endocrinology 145, 3495, 2004
大鼠SD大鼠,雄性 150-220 克Dorsal root ganglion neuronsCollagenase Type 4: 0.125%DMEM/Ham's F12Hu Hong-Zhen, Gu Qihai, Wang Chunbo, Colton Craig K, Tang Jisen, Kinoshita-Kawada Mariko, Lee Lu-Yuan, Wood Jackie D, Zhu Michael X:2-aminoethoxydiphenyl borate is a common activator of TRPV1, TRPV2, and TRPV3, J Biol Chem 279, 35741-8, 2004
Trypsin: 0.05%
大鼠大鼠, 15 周NeuronsPapain: 0.2%Hibernate AEvans J, Sumners C, Moore J, Huenman MJ, Deng J, Gelband CH, and Shaw G: Characterization of mitotic neurons derived from adult rat hypothalamus and brain stem, J Neurophysiol 87, 1076, 2002
大鼠SD大鼠,雄性, 270-330 克Spinal progenitor cellsPDS kit: with modificationsNeurobasal mediumLin CR, Wu PC, Shih HC, Cheng JT, Lu CY, Chou AK, Yang LC.:Intrathecal spinal progenitor cell transplantation for the treatment of neuropathic pain, Cell Transplant 11, 17, 2002
大鼠大鼠, 食魚貂, 8-9 周Adult progenitorPapain: 2.5 u/mlDMEM/F-12Lie DC, Dziewczapolski G, Willhoite AR, Kaspar BK, Shults CW, Gage FH.: The adult substantia nigra contains progenitor cells with neurogenic potential, J Neurosci 22, 6639, 2002
Deoxyribonuclease I: 250 u/ml
Neutral Protease: 1 u/ml
大鼠SD大鼠, 胚胎Sciatic Nerve and Gut Neural Crest Stem CellsCollagenase Type 4: 0.025%HBSSBixby, S., Kruger, G., Mosher, J., Joseph, N. and Morrison, S.: Cell-Intrinsic Differences Between Stem Cells from Different Regions of the Peripheral Nervous System Regulate the Generation of Neural Diversity.,Neuron 35, 643, 2002
Trypsin: 0.005%
Deoxyribonuclease I: 0.05%
大鼠大鼠Neurons, hippocampalPapain: 15 - 20 u/mlEagle's MEMLiu, Q., Kawai, H., and Berg, D.: B-Amyloid Peptide Blocks the Response ofa7-Containing Nicotinic Receptors on Hippocampal Neurons, PNAS 98 (8), 4734, 2001
(see reference)
大鼠大鼠, 新生的, 7 天 大NeuronsTrypsin: 0.25%MEM10Acosta, C., Fabrega, A., Masco, D., and Lopez, H.: A Sensory Neuron Subpopulation with Unique Sequential Survival Dependence on Nerve Growth Factor and Basic Fibroblast Growth Factor during Development, J Neurosci 21 (22), 8873, 2001
大鼠大鼠, 胚胎18-19 天Hippocampal neuronsPDS kit: see referenceDMEMMabuchi T, Kitagawa K, Kuwabara K, Takasawa K, Ohtsuki T, Xia Z, Storm D, Yanagihara T, Hori M, Matsumoto M: Phosphorylation of cAMP response element-binding protein in hippocampal neurons as a protective response after exposure to glutamate in vitro and ischemia in vivo, J Neurosci 21, 9204-13, 2001
大鼠大鼠, 胚胎, 天 18CorticalPapain: Neurobasal medium and DMEMO'Connor, S., Andreadis, J., Shaffer, K., Ma, W., Pancrazio, J., and Stenger, D.: Immobilization of Neural Cells in Three-Dimensional Matrices for Biosensor Applications, Biosensors & Bioelectronics 14, 871, 2000
大鼠大鼠, Wister, postnatal, P1-3 天Neurons, hippocampalPapain: 20 u/mlEBSSNeuhoff, H., Roeper, J., Schweizer, M.: Activity-Dependent Formation of Perforated Synapses in Cultured Hippocampal Neurons, Eur J Neurosci 11, 4241, 1999
大鼠SD大鼠, 雌性, pre克nantSciatic nervesTrypsin: 0.025%L-15 mediumMorrison, S., White, P., Zock, C., and Anderson, D.: Prospective Identification, Isolation by Flow Cytometry, and In Vivo Self-Renewal of Multipotent Mammalian Neural Crest Stem Cells, Cell 96, 737, 1999
Collagenase Type 3: 0.1%(see reference)
大鼠大鼠Spinal cordTrypsin: 0.133%HBSS and PIPESJohansson, C., Momma, S., Clarke, D., Risling, M., Lendahl, U., and Frisen, J: Identification of a Neural Stem Cell in the Adult Mammalian Central Nervous System, Cell 96, 25, 1999
(包括 小鼠)
大鼠SD大鼠, 胚胎Sciatic nervesTrypsin: 0.025%L-15 mediumMorrison, S., White, P., Zock, C., and Anderson, D.: Prospective Identification, Isolation by Flow Cytometry, and In Vivo Self-Renewal of Multipotent Mammalian Neural Crest Stem Cells, Cell 96, 737, 1999
Collagenase Type 3: 0.1%(see reference)
大鼠大鼠, 小白鼠), 2-5 天 大Neurons, hippocampalPapain: 20 u/mlEBSSWilding, T., and Huettner, J.: Activation and Desensitization of Hippocampal Kainate Receptors, J Neurosci 17 (8), 2713, 1997
大鼠SD大鼠和Wistar大鼠, 新生的, 0-21 天, 兩者之間任意一個Myenteric gangliaTrypsin: 0.05%MEM-HEPESSchafer, K., Saffrey, M., Burnstock, G., and Mastres-Ventura, P.: A New Method for the Isolation of Myenteric Plexus from the Newborn Rat Gastrointestinal Tract, Brain Res Proto 1, 109, 1997
大鼠大鼠, 幼崽, 24-48 小時大Neurons, hippocampalPapain: 20 u/mlHarvest bufferHall, R., and Soderling, T.: Differential Surface Expression and Phosphorylation of the N-Methyl-D-Aspartate Receptor NR1 and NR2 in Cultured Hippocampal Neurons, J Biol Chem 272 (7), 4135, 1997
大鼠SD大鼠, 成年的, 250-300克, P8Neurons, DRGNeutral Protease: 0.5%L-15 w/ CO2Davies, S., Fitch, M., Memberg, S., Hall, A, Raisman, G., and Silver, J.:Regeneration of Adult Axons in White Matter Tracts of the Central Nervous System, Nature 390, 680, 1997
大鼠大鼠, 1-4 天 大Neurons, hippocampalPapain: 20 u/mlMEMTwitchell, W., Brown, S., and Mackie, K.: Cannabinoids Inhibit N- and P/Q-Type Calcium Channels in Cultured Rat Hippocampal Neurons, J Neurophysiol 78, 43, 1997
大鼠SD大鼠, 食魚貂Hippocampal neuronsPapain: 0.2%HibernateA/B27Brewer, G.J.: Isolation and culture of adult rat hippocampal neurons of any age, J Neurosci Methods 71, 143, 1997
大鼠大鼠, E18Hippocampal neuronsPapain: 20 u/mlMEMLiu QY, Schaffner AE, Li YX, Dunlap V, Barker JL: Upregulation of GABAA current by astrocytes in cultured 胚胎rat hippocampal neurons, J Neurosci 16, 2912-23, 1996
Deoxyribonuclease I: 0.01%
大鼠大鼠, 1-6 天DRG neuronsPapain: 20 u/mlHBSSRobertson, S., Rae, M., Rowan, E. and Kennedy, C.: Characterization of a P2X-Purinoceptor in Cultured Neurones of the Rat Dorsal Root Ganglia., Br J Pharmacol 118, 951, 1996
Collagenase Type 1: 150 u/ml
Neutral Protease: 0.8%
大鼠大鼠, 幼崽, 14 天 大Neurons, sympatheticNeutral Protease: 0.24%HBSSMcFarlane, S., and Cooper, E.: Extrinsic Factors Influence the Expression of Voltage-Gated K Currents on Neonatal Rat Sympathetic Neurons, J Neurosci 13 (6), 2591, 1993
大鼠,SD大鼠, 11-14 天 大, 29-32 克Basal forebrain neuronsTrypsin: 0.125%Gey's BSSAllen, T., Sim, J., and Brown, D.: The Whole-Cell Calcium Current in Acuy Dissociated Magnocellular Cholinergic Basal Forebrain Neurones of the Rat,J Physiol 460, 91, 1993
大鼠Hanover Wistar大鼠, 7-12 天Postnatal dopamine neuronsTrypsin: 0.035%(see reference)Rayport, S., Sulzer, D., Shi, W., Sawasdikosol, S., Monaco, J., Batson, D., and Rajendran, G.: Identified Postnatal Mesolimbic Dopamine Neurons in Culture: Morphology and Electrophysiology, J Neurosci 12 (11), 4264, 1992
大鼠SD大鼠, 18-天-大, 胎兒HippocampalTrypsin: 0.2%HBSSCheng, B., and Mattson, M.: IGF-I and IGF-II Protect Cultured Hippocampal and Septal Neurons Against Calcium-Mediated Hypoglycemic Damage, J Neurosci 12 (4), 1558, 1992
大鼠大鼠Stem, neural crestCollagenase: 0.075%Ringer's solutionStemple, D., and Anderson, D.: Isolation of a Stem Cell for Neurons and Glia from the Mammalian Neural Crest, Cell 71, 973, 1992
大鼠SD大鼠, 懷孕的, 查爾斯河Pyramidal neuronsTrypsin: 0.027%HEPESBuchhalter, J., and Dichter, M.: Electrophysiological Comparison of Pyramidal and Slate Nonpyramidal Neurons in Dissociated Cell Culture of Rat Hippocampus, Brain Res Bull 26, 333, 1991
Nonpyramidal neurons
大鼠SD大鼠, 胚胎, 19-21 天 妊娠期Schwann, dorsal root gangliaTrypsin: 0.25%HBSS, CMFMithen, F., Reiker, M., and Birchem, R.: Effects of Ethanol on Rat Schwann Cell Proliferation and Myelination in Culture, In Vitro Cell Dev Biol 26, 129, 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 Bovine Adrenal Chromaffin Paraneurons, In Vitro Cell Dev Biol 26, 335, 1990
(包括 牛)Adrenal chromaffin 
 Paraneurons
大鼠SD大鼠, 8 天 大 幼崽Cerebellar neuronsTrypsin: 0.025%Eagle's MEMNovelli, A., Reilly, J., Lysko, P., and Henneberry, R.: Glutamate Becomes Neurotoxic Via the N-methyl-D-aspartate Receptor When Intracellular Energy Levels are Reduced, Brain Res 451, 205, 1988
大鼠大鼠,胎兒, 18 天 大Hippocampal neuronsTrypsin: 0.2%Eagle's MEMMattson, M., and Kater, S.: Isolated Hippocampal Neurons in Cryopreserved Long-Term Cultures: Development of Neuroarchitecture and Sensitivity to Nmda, Int J Dev Neurosci 6 (5), 439, 1988
大鼠大鼠, 產(chǎn)后Septal neuronsPapain: 0.05%PBS, CMFHatanaka,H., Tsukui, H., Nihonmatsu, I.: Septal Cholinergic Neurons From Postnatal Rat Can Survive In The Dissociate Culture Conditions In The Presence Of Nerve Growth Factor, Neurosci Lett 79, 85, 1987
大鼠大鼠, 小白鼠, 1-15 天Neurons, visual cortexPapain: 20 u/mlBSSHuettner, J., and Baughman, R.: Primary Culture of Identified Neurons From the Visual Cortex of Postnatal Rats, J Neurosci 6, 3044, 1986
(see reference)
大鼠SD大鼠, 雌性, 4 月 大CNS cellsTrypsin: 0.25%EBSSWood, P., and Bunge, R.: Evidence That Axons are Mitogenic for Oligodendrocytes Isolated From Adult Animals, Nature 320, 756, 1986
大鼠大鼠, 胚胎, 15 天Dorsal horn neuronsTrypsin: 0.025%Ham's F-12Jahr, C. and Jessell, T.: Synaptic Transmission between Dorsal Root Ganglion and Dorsal Horn Neurons in Culture: Antagonism of Monosynaptic Excitatory Postsynaptic Potentials and Glutamate Excitation by Kynurenate,J Neurosci 5, 2281, 1985
Spinal
大鼠大鼠, 胎兒, 18-20 天Hippocampal neuronsTrypsin: 0.25%HBSS, CMFBartlett, W. and Banker, G.: An Electron Microscopic Study of the Development of Axons and Dendrites by Hippocampal Neurons in Culture. I. Cells Which Develop Without Intercellular Contacts, J Neurosci 4, 1944, 1984
大鼠大鼠, 產(chǎn)后Ganglion, retinaPapain: 12.5 u/mlHBSS w/5 mM HEPESLeifer, D., Lipton, S., Barnstable, C., and Masland, R.: Monoclonal Antibody to Thy-1 Enhances Regeneration of Processes by Rat Retinal Ganglion Cells in Culture, Science 224, 303, 1984
大鼠SD大鼠, 幼崽RetinaTrypsin: 0.25%Ham's F-12Sarthy PV, Curtis BM, and Catterall WA.: Retrograde Labeling, Enrichment, and Characerization of Retinal Ganglion Cells from the Neonatal Rat, J Neurosci 3 (12), 2532, 1983
大鼠大鼠, W/FU, 5-8 天Neurons and glialTrypsin: 0.25%MEMRaff, M., Fields, K., Hakomori, S., Mirsky, R., Pruss, R., and Winter, J.:Cell-Type-Specific Markers for Distinguishing and Studying Neurons And the Major Classes of Glial Cells in Culture, Brain Res 174, 283, 1979
(see reference)
大鼠大鼠, 胎兒Neurons, sympatheticTrypsin: 0.25%L-15 or HBSS, CMFWakshull, E., Johnson, M., Burton, H.: Postnatal Rat Sympathetic Neurons In Culture. 1. A Comparison With 胚胎Neurons, J Neurophysiol 42, 1410, 1979
大鼠 Wistar/Furth大鼠, 新生的SchwannTrypsin: 0.25%DMEMBrockes, J., Fields, K., and Raff, M.: Studies on Cultured Rat Schwann Cells. I. Establishment of Purified Populations From Cultures of Peripheral Nerve,Brain Res 165, 105, 1979
(包括 牛)
大鼠大鼠, 胚胎Neurons, corticalTrypsin: 0.027%MEMDichter, M.: Rat Cortical Neurons in Cell Culture: Culture Methods, Cell Morphology, Electrophysiology, and Synapse Formation, Brain Res 149, 279, 1978
大鼠大鼠, 新生的Neurons, sympatheticCollagenase Type 1: 0.01%Hank's solution, CFReichardt, L., Patterson, P.: Neurotransmitter Synthesis and Uptake by Isolated Sympathetic Neurones in Microcultures, Nature 270, 147, 1977
大鼠大鼠, 新生的Neurons, superior cervical gangliaTrypsin: 0.1%Basal L-15 mediumMains, R., and Patterson, P.: Primary Cultures of Dissociated Sympathetic Neurons I. Establishment of Long-Term Growth in Culture and Studies of Differentiated Properties, J Cell Biol 59, 329, 1973
蠑螈蠑螈, 18-25 cmRetinaPapain: 14 u/mlSalineTownes-Anderson, E., MacLeish, P., and Raviola, E.: Rod Cells Dissociated from Mature 蠑螈 Retina: Ultrastruture and Uptake of Horseradish Peroxidase, J Cell Biol 100, 175, 1985
蠑螈蠑螈 (A.tigrinum)Photoreceptors, retinaPapain: 0.05%(see reference)Bader, C., MacLeish, P., and Schwartz, E.: Responses to Light of Solitary Rod Photoreceptors Isolated From Tiger 蠑螈 Retina, Proc Natl Acad Sci U S A 75, 3507, 1978
貝類動物海兔NeuronsNeutral Protease: 1.0%L-15-ASWLee, A., Decourt, B. and Suter, D.: Neuronal Cell Cultures from Aplysia for High-Resolution Imaging of Growth Cones., J Vis Exp 12, 662, 2008
貝類動物蝸牛(旋節(jié)螺), 白化病的, 成年的Buccal ganglia; SLT muscleTrypsin: 0.2%DMEMZoran, M., Doyle, R. and Haydon, P.: Target Contact Regulates the Calcium Responsiveness of the Secretory Machinery During Synaptogenesis,Neuron 6, 145, 1991
貝類動物田螺(旋節(jié)螺) 白化病的, 成年的Somata, buccal gangliaTrypsin: 0.2%Antibiotic saline, Leibowitz 50%Haydon, P.: The Formation of Chemical Synapses Between Cell-Cultured Neuronal Somata, J Neurosci 8, 1032, 1988
貝類動物旋節(jié)螺Buccal gangliaTrypsin: 0.2%L-15 mediumCohan, C., Haydon, P., and Kater, S.: Single Channel Activity Differs in Growing and Nongrowing Growth Cones of Isolated Identified Neurons of Helisoma, J Neurosci Res 13, 285, 1985
貝類動物軟體動物, 青年的 1-2 克 或者成年的50-100 克NeuronsProtease: 1%L15 mediumSchacher, S., and Proshanksy, E.: Neurite Regeneration by Aplysia Neurons in Dissociated Cell Culture: Modulation by Aplysia Hemolymph and the Presence of the Initial Axonal Segment, J Neurosci 3 (12), 2403, 1983
LUQ cells
貝類動物海兔LUQ cellsProtease: 1%L15 mediumCamardo, J., Proshansky, E., and Schacher, S.: Identified Aplysia Neurons Form Specific Chemical Synapses in Culture, J Neurosci 3 (12), 2614, 1983
RUQ cells
龜龜 (紅耳龜)RetinalPapain: 0.1% (13.5 u/mg)Kreb's RingerLam, D.: Biosynthesis of Acetylcholine in Turtle Photoreceptors, Proc Natl Acad Sci U S A 69, 1987, 1972

 

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