神經(jīng)組織消化手冊(cè)

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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(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(duì)其組織消化和細(xì)胞收集的作用機(jī)制卻沒(méi)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(guò)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(lèi)型

2、物種來(lái)源

3、動(dòng)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(méi)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(guò)查閱文獻(xiàn)來(lái)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(lái)源于組織胞外基質(zhì)的復(fù)雜而動(dòng)態(tài)的特性，以及過(guò)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(lèi)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(shí)和方法，描述了標(biāo)準(zhǔn)的實(shí)驗(yàn)步驟，以便于制定一個(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(lèi)型和物種分類(lèi)，詳見(jiàn)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(dòng)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(lèi)型的消化。

神經(jīng)組織消化手冊(cè)

物種	詳細(xì)來(lái)源	細(xì)胞名稱(chēng)	消化用酶	培養(yǎng)基	參考文獻(xiàn)
鳥(niǎo)類(lèi)	小雞, 白來(lái)杭雞, 胚胎	Cilary ganglion neurons	Collagenase Type 1: 0.1%	PBS	Temburni, 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	Heart	Trypsin: 0.06%	25mM HEPES buffered Locke's solution, CMF	Trifaro, 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 endothelial	Collagenase/Dispase: 0.1%	MEM	Bowman, 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	Oligodendroglia	Trypsin: 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(lái)杭雞, 胚胎, 17-21 天	Neurons	Papain: 40 u/ml	HEPES	Raman, 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, retina	Trypsin: 0.1%	Tyrode's solution, CMF	Moyer, 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(lái)杭雞, 胚胎	Cerebral neurons	Trypsin: Trypsin: 0.125-0.25%	DMEM	Coates, 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(lái)杭雞, 胚胎	Spinal cord	Trypsin: 0.05%	Phosphate buffer	Schnaar, 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 neurons	Trypsin: 0.1%	Puck's saline, CMF	Choi, 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(lái)杭雞胚胎, 8 天	Neurons, ganglia	Trypsin: 0.25%	HBSS, CMF	Bottenstein, 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(lái)杭雞, 種蛋	Ciliary ganglion neurons	Trypsin: 0.25%	Eagle's MEM	Tuttle, 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 neurons	Collagenase: 0.01%	Eagle's MEM	Mudge, 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 ganglia	Trypsin: 0.25%	Krebs Phosphosaline	McCarthy, 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(lái)杭雞, 胚胎, 7 天	Neural retina	Collagenase: 0.25%	HBSS	Wiseman, 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(yú)	魚(yú), 斑馬魚(yú)	Motor neurons	Collagenase Type 2: 0.1%	See reference	Sakowski, 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(yú)	Dania rerio斑馬魚(yú), 胚胎	Rohon-Beard neurons	Trypsin: 0.2%	L-15/Hepes	Won, 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(yú)	Dania rerio斑馬魚(yú), 胚胎	Neurons	PDS kit: with modifications	EBSS	Cerda, 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(yú)	黑色的魅影kni克e魚(yú), 成年的(黑魔鬼魚(yú))	Neurons, spinal cord	Trypsin: 0.4%	PBS, CMF	Anderson, 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	Neuron	Collagenase Type 1: 0.1%	Steinberg's solution	Takahashi, 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, enteric	Trypsin: 0.125%	Medium 199	Jessen, 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 neurons	Collagenase Type 1: 0.02%	PBS	Liman 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(lèi)	人類(lèi)	Neural, various	Papain: 12 u/ml	see reference	Panchision 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(lèi)	人類(lèi), 9 個(gè)月	Neurons	Collagenase Type 4: 1.33%	DMEM/F12	Dietrich 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(lèi)	人類(lèi), 26 周	Neural progenitor cells	Papain: 2.5 u/ml	DMEM/F-12	Fuja 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(lèi)	人類(lèi), 成年人	Ventricular epithelial	Papain: 11.4 u/ml	DMEM/F12	Roy 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(lèi)	人類(lèi), 5-65 歲	Retinal pigment epithelial (RPE)	Trypsin: 0.25%	HBSS	Von 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(lèi)	人類(lèi)	Dorsal root ganglion neurons	Collagenase Type 1: 0.2%	DMEM/F-12	Dib-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(chóng)	果蠅, 幼蟲(chóng)	Neuroblasts	Collagenase Type 1: 0.1%	Rinaldini solution	Berger, 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(chóng)	果蠅	Dendrites	Collagenase: 0.05%	HBSS	Sanchez-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(chóng)	雙斑蟋	Giant interneurons	Collagenase: 0.05%	Leibovitz's L15	Kloppenburg, 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 neurons	Collagenase Type 2: 0.1%	DMEM/F-12	Lee, 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 cells	Collagenase: 0.05-0.1%	DMEM	Stettner, 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	Oligodendrocytes	Papain: 0.1%	HBSS	Harris, 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 neurons	PDS kit: per instructions	DMEM	Williams, 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	Neurosperes	Papain: 0.1%	NeuroCult NSC	Stuart, 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	Schwann	Collagenase Type 1: 0.01%	DMEM	Masaki, 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 cells	Papain: 0.09%	DMEM	Medina-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 precursors	Trypsin: 0.13%	see reference	Babona-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 neurons	Papain: 10-20 ul	HBSS	Goetz, J. and Trimarchi, J.: Single-Cell Profiling of Developing and Mature Retinal Neurons., J Vis Exp 62, e3824, 2012
小鼠	小鼠	Neurons	Papain: 100 ul	DMEM	Li, 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, P4-5	Neuroshere	Trypsin: 0.25%	Pro-N	Ziegler, 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 cells	Papain: 0.01%	DMEM/F12	Bracko, 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 neurons	Papain: 0.05%	PBS/DMEM	Pollari, 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 ganglia	Papain: 0.15%	DMEM	O'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	Motorneurons	Trypsin: 0.025%	HBSS	Conrad, 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 ensheathing	Collagenase: 0.15%	DMEM	Radtke, 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 Precursor	Papain: 2.5 u/ml	Neurobasal A	Babu, 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 neurons	Papain: 1%	HBSS	Brown, 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 leukocytes	Collagenase Type 4: 300 u/ml	HBSS	Sayed, 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 microganglia	Papain: 0.2%	Hibernate A	Yip, 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 neurons	Trypsin: 0.1%	DMEM	Radad 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 neurons	Collagenase: 0.2%	Ham's F12	Pedrola, 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(gè)月	Neurons, neurospheres	Papain: 0.2%	Hibernate	Brewer Gregory J, Torricelli John R: Isolation and culture of adult neurons and neurospheres, Nat Protoc 2, 1490-8, 2007
小鼠	小鼠	Neural, various	Papain: 12 u/ml	see reference	Panchision 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, DRG	Papain: 20 u/ml	HBSS	Malin, 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	Neurons	Papain: 0.2%	DMEM	Eide, 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 cells	Trypsin: 0.25%	PBS	Gonzalez 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	Neurons	PDS kit: per instructions	EBSS	Gill 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, C57BL	Cerebellar granule cell precursors	Papain: 16.5 u/ml	Dulbecco's PBS	Okano-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, ganglia	Papain: 20 u/ml	HBSS	Savchenko 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 neurons	Papain: 20 u/ml	HEPES buffered saline	Roberts, 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	Neurons	Trypsin: 0.25%	NGF-containing medium	Deshmukh, 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 SCG	Trypsin: 0.25%	L-15 medium	Lee, 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 precursors	Trypsin: 0.1%	DMEM	Richards 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, neonate	Neurons, dorsal root ganglion	Trypsin: 0.25%	HBSS	Quinn, 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	Precursor	Trypsin: 0.5%	PBS	Kitani, 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s	Papillae, taste receptor	Pronase E: 0.15%	Carbonate-Phosphate buffer	Spielman, 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 neurons	Trypsin: 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(shí)驗(yàn)室老鼠), 成年的	Neurons, spinal cord	Collagenase Type 3: 0.25%	Hank's BSS, CMF	Eagleson, 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	Neural	Trypsin NF 1:250: 50 0.25%	BSS	Shrier, B., Wilson, S., and Nirenberg, M.: Cultured Cell Systems and Methods for Neurobiology, Vol. 32,, , 765, 1974
綿羊	羊羔	Oligodendroglia	Trypsin: 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 ganglia	Papain: 2 u/ml	HBSS	Si 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 crest	Trypsin: 0.05%	MEM, HBSS	Sieber-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, DRG	PDS kit: per instructions	Neurobasal	Rheaume, 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 neurons	Papain: 2 u/ml	DMEM-PBS	Oyanagi, 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 Ganglia	Collagenase Type 3: 0.2%	HBSS	Murayama, 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 neurons	Collagenase Type 2: 0.15%	HBSS	Yu, 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 cells	Collagenase: 0.05-0.1%	DMEM	Stettner, 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 Neurons	Papain: 25 u/ml	L-15	Thurner, 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 neuronal	PDS kit: per instructions	Neurobasal A	Loktev, 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 ganglia	Papain: 20 u/ml	HBSS	Yan, 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 P1-3	Sympathetic neurons	Collagenase Type 2: 0.1%	DMEM/F-12	Ghogha, 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 Ganglion	Papain: 20 u/ml	L-15	Bosmans, 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, fibroblast	Collagenase: 0.125%	DMEM	East, 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(yú)貂, 7-21 月	Hippocampal neurons	Papain: 0.2%	Hibernate A	Chen, 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, E18	Neuronal	Papain: 0.2%	Neurobasal E	Peltier, 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 neurons	PDS kit: per instructions	PBS	Tanaka, 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 18	Hippocampal neurons	Papain: 20 u/ml	Neurobasal/B27	Liu, 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 glias	Papain: 0.5 u/ml	DMEM	Yokosuka 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 ganglion	Collagenase: 0.05%	L-15	Sakisaka, 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 neurons	Papain: 20 u/ml	PBS	Frank, 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 neurons	PDS kit: per instructions	Neurobasal A	Obradovic 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	astrocytes	PDS kit: per instructions	DMEM	Lacroix-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 reference	Neurobasal A	Mothe, 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, E19	Dorsal root ganglia	PDS kit: per instructions	MEM/Ham's F12	Gavva 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, E15	Vomeronasal receptor neurons	Collagenase/Dispase: 0.1%	DMEM/F12	Moriya-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 astrocytes	Papain: see reference	DMEM	Floyd 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 ganglion	Collagenase Type 4: 20 u/ml	DMEM	Pedraza 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 neurons	Papain: 20 u/ml	CMF Hanks	Connor, 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 neurons	PDS kit: per instructions	DMEM	Lovshin 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 neurons	Collagenase Type 4: 0.125%	DMEM/Ham's F12	Hu 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	Neurons	Papain: 0.2%	Hibernate A	Evans 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 cells	PDS kit: with modifications	Neurobasal medium	Lin 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(yú)貂, 8-9 周	Adult progenitor	Papain: 2.5 u/ml	DMEM/F-12	Lie 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 Cells	Collagenase Type 4: 0.025%	HBSS	Bixby, 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, hippocampal	Papain: 15 - 20 u/ml	Eagle's MEM	Liu, 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	Neurons	Trypsin: 0.25%	MEM10	Acosta, 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 neurons	PDS kit: see reference	DMEM	Mabuchi 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 18	Cortical	Papain:	Neurobasal medium and DMEM	O'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, hippocampal	Papain: 20 u/ml	EBSS	Neuhoff, 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nant	Sciatic nerves	Trypsin: 0.025%	L-15 medium	Morrison, 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 cord	Trypsin: 0.133%	HBSS and PIPES	Johansson, 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 nerves	Trypsin: 0.025%	L-15 medium	Morrison, 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, hippocampal	Papain: 20 u/ml	EBSS	Wilding, 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(gè)	Myenteric ganglia	Trypsin: 0.05%	MEM-HEPES	Schafer, 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(shí)大	Neurons, hippocampal	Papain: 20 u/ml	Harvest buffer	Hall, 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, P8	Neurons, DRG	Neutral Protease: 0.5%	L-15 w/ CO₂	Davies, 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, hippocampal	Papain: 20 u/ml	MEM	Twitchell, 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(yú)貂	Hippocampal neurons	Papain: 0.2%	HibernateA/B27	Brewer, G.J.: Isolation and culture of adult rat hippocampal neurons of any age, J Neurosci Methods 71, 143, 1997
大鼠	大鼠, E18	Hippocampal neurons	Papain: 20 u/ml	MEM	Liu 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 neurons	Papain: 20 u/ml	HBSS	Robertson, 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, sympathetic	Neutral Protease: 0.24%	HBSS	McFarlane, 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 neurons	Trypsin: 0.125%	Gey's BSS	Allen, 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 neurons	Trypsin: 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	Hippocampal	Trypsin: 0.2%	HBSS	Cheng, 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 crest	Collagenase: 0.075%	Ringer's solution	Stemple, 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 neurons	Trypsin: 0.027%	HEPES	Buchhalter, 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 ganglia	Trypsin: 0.25%	HBSS, CMF	Mithen, 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	Heart	Trypsin: 0.06%	25mM HEPES buffered Locke's solution, CMF	Trifaro, 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 neurons	Trypsin: 0.025%	Eagle's MEM	Novelli, 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 neurons	Trypsin: 0.2%	Eagle's MEM	Mattson, 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 neurons	Papain: 0.05%	PBS, CMF	Hatanaka,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 cortex	Papain: 20 u/ml	BSS	Huettner, 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 cells	Trypsin: 0.25%	EBSS	Wood, 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 neurons	Trypsin: 0.025%	Ham's F-12	Jahr, 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 neurons	Trypsin: 0.25%	HBSS, CMF	Bartlett, 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, retina	Papain: 12.5 u/ml	HBSS w/5 mM HEPES	Leifer, 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	Retina	Trypsin: 0.25%	Ham's F-12	Sarthy 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 glial	Trypsin: 0.25%	MEM	Raff, 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, sympathetic	Trypsin: 0.25%	L-15 or HBSS, CMF	Wakshull, 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	Schwann	Trypsin: 0.25%	DMEM	Brockes, 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, cortical	Trypsin: 0.027%	MEM	Dichter, 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, sympathetic	Collagenase Type 1: 0.01%	Hank's solution, CF	Reichardt, 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 ganglia	Trypsin: 0.1%	Basal L-15 medium	Mains, 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 cm	Retina	Papain: 14 u/ml	Saline	Townes-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, retina	Papain: 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(lèi)動(dòng)物	海兔	Neurons	Neutral Protease: 1.0%	L-15-ASW	Lee, 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(lèi)動(dòng)物	蝸牛(旋節(jié)螺), 白化病的, 成年的	Buccal ganglia; SLT muscle	Trypsin: 0.2%	DMEM	Zoran, M., Doyle, R. and Haydon, P.: Target Contact Regulates the Calcium Responsiveness of the Secretory Machinery During Synaptogenesis,Neuron 6, 145, 1991
貝類(lèi)動(dòng)物	田螺(旋節(jié)螺) 白化病的, 成年的	Somata, buccal ganglia	Trypsin: 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(lèi)動(dòng)物	旋節(jié)螺	Buccal ganglia	Trypsin: 0.2%	L-15 medium	Cohan, 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(lèi)動(dòng)物	軟體動(dòng)物, 青年的 1-2 克或者成年的50-100 克	Neurons	Protease: 1%	L15 medium	Schacher, 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(lèi)動(dòng)物	海兔	LUQ cells	Protease: 1%	L15 medium	Camardo, 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)	Retinal	Papain: 0.1% (13.5 u/mg)	Kreb's Ringer	Lam, D.: Biosynthesis of Acetylcholine in Turtle Photoreceptors, Proc Natl Acad Sci U S A 69, 1987, 1972

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