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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 | 結腸組織 | 內(nèi)皮組織 | 上皮組織 |
眼 | 心臟 | 腸 | 腎 |
肝 | 肺 | 淋巴結 | 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 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 | 魚, 斑馬魚 | 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 | Dania rerio斑馬魚, 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 | Dania rerio斑馬魚, 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 | 黑色的魅影kni克e魚, 成年的(黑魔鬼魚) | 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 | 人類 | 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 | 人類, 9 個月 | 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 | 人類, 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 | 人類, 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 | 人類, 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 | 人類 | 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 | 果蠅, 幼蟲 | 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 | 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 | 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 (白變種實驗室老鼠), 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 | 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 |
| 貝類動物 | 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 |
| 貝類動物 | 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 |
| 貝類動物 | 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 |
| 貝類動物 | 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 |
| 貝類動物 | 軟體動物, 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 | |||||
| 貝類動物 | 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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