原代细胞分离中，针对脑组织涉及到微血管内皮、脑内皮细胞、神经元和小胶质细胞、微血管等等的解离通常会使用酶，如何选择合适的酶来完成对应脑组织的解离，以及原代细胞采集的常规操作， 艾美捷针对脑相关的组织分类解离分种属、不同的细胞、酶组合推荐，推荐浓度、培养基以及参考文献，详细汇总指南：
本技术指南描述了标准实验室程序； 为建立细胞分离方案提供了一种合乎逻辑的实验方法； 并列出了许多组织特异性参考资料，以帮助开发有效的方法。更多其他组织的解离指南，点击查看：各组织分离指南。
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| 种属 | 种属详情 | 细胞名称 | 酶组合推荐，含推荐浓度 | 培养基 | 参考文献 |
| 牛 | Bovine | 微血管内皮 / Microvascular endothelial | Neutral Protease: 0.005% | Medium 199 | Kanda, T., Yoshino, H., Ariga, T., Yamawaki, M., and Yu, R.: Glycosphingolipid Antigens in Cultured Microvasular Bovine Brain Endothelial Cells: Sulfoglucuronosyl Paragloboside as a Target of Monoclonal IgM in Demyelinative Neuropathy, J Cell Biol 126 (1), 235, 1994 |
| 牛 | Bovine | 脑内皮细胞 / Brain endothelial cells | Collagenase Type 2: 0.35% | DMEM | Wolburg H, Neuhaus J, Kniesel U, Krauss B, Schmid EM, Ocalan M, Farrell C, Risau W: Modulation of tight junction structure in blood-brain barrier endothelial cells. Effects of tissue culture, second messengers and cocultured astrocytes, J Cell Sci 107, 1347, 1994 |
| 牛 | Bovine | 内皮 / Endothelial | Neutral Protease: 0.125% | MEM | Miller, D., Audus, K., and Borchardt, R.: Application of Cultured Endothelial Cells of the Brain Microvasculature in the Study of the Blood-Brain Barrier, J Tiss Cul Meth 14, 217, 1992 |
| 牛 | Bovine, adult | 脑动脉，内皮 / Cerebral artery，Endothelial | Collagenase: 0.2% | HBSS | Machi, T., Kassell, N.F., and Scheld, W.M.: Isolation and Characterization of Endothelial Cells From Bovine Cerebral Arteries, In Vitro Cell Dev Biol 26, 291, 1990 |
| 牛 | Bovine | 毛细血管内皮 / Capillary endothelial | Collagenase: 0.1% | DMEM | Estrada, C., Bready, J., Berliner, J., and Cancilla, P.: Choline Uptake by Cerebral Capillary Endothelial Cells in Culture, J Neurochem 54, 1467, 1990 |
| 牛 | Bovine | 内皮 / Endothelial | Neutral Protease: 0.5% | MEM | Audus, K., and Borchardt, R.: Characterization of an In Vitro Blood-Brain Barrier Model System for Studying Drug Transport and Metabolism, Pharm Res 3 (2), 81, 1986 |
| 牛 | Bovine | 内皮，脑动脉 / Endothelial，Brain arteries | Collagenase Type 2: 0.2% | Dulbecco's PBS | Goetz, I.., Warren, J., Estrada, C., Roberts, E., and Krause, D.: Long-Term Cultivation of Arterial and Capillary Endothelium From Adult Bovine Brain, In Vitro Cell Dev Biol 21, 172, 1985 |
| 牛 | Calf | 少突胶质细胞，神经 / Oligodendroglia，Neural | 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 |
| 豚鼠 | Guinea-pig, 200-400 g | 神经元 / Neurons | Trypsin: 0.06-0.08% | PIPES saline | Kay, A.R., and Wong, R.K.S.: Isolation of Neurons Suitable for Patch-Clamping from Adult Mammalian Central Nervous Systems, J Neurosci Methods 16, 227, 1986 |
| 人 | Human | 小胶质细胞 / Microglia | Collagenase Type 1: 300 u/ml； Trypsin: 0.125% |
DMEM | Mizee, M., Miedema, S., van der Poel, M., Adelia , S., van Strien, M., Melief, J., Smolders, J., Hendrickx, D., Heutinck, K., Hamann, J. and Huitinga, I.: Isolation of Primary Microglia from the Human Post-Mortem Brain: Effects of Ante- and Post-Mortem Variables., Acta Neuropathol 5, 16, 2017 |
| 人 | Human, adult | 神经元 / Neuronal | Papain: 20 u/ml | Neurobasal | Spaethling, J., Na, Y., Lee, J., Ulyanova, A., Baltuch, G., Bell, T., Brem, S., Chen, H., Dueck, H., Fisher, S., Garcia, M., Khaladkar, M., Kung, D., Lucas, T., O'Rourke, D. and Stefanik, D.: Primary Cell Culture of Live Neurosurgically Resected Aged Adult Human Brain Cells and Single Cell Transcriptomics., Cell Rep 18, 791-803, 2017 |
| 人 | Human, fetal and mature | 星形胶质细胞和神经元 / Astrocytes and neurons | Papain: 7.5-20 u/ml | RPMI | Zhang, Y., Sloan, S., Clarke, L., Caneda, C., Plaza, C., Blumenthal, P., Vogel, H., Steinberg, G., Edwards, M., Li, G., Duncan, J., Cheshier, S., Shuer, L., Chang, E., Grant, G., Gephart, M. and Barres, B.: Purification and Characterization of Progenitor and Mature Human Astrocytes Reveals Transcriptional and Functional Differences with Mouse., Neuron 89, 37-53, 2016 |
| 人 | Human | 小胶质细胞 / Microglia | Neutral Protease: 10 u/ml； Papain: 2.5 u/ml |
Hibernate A | Rustenhoven, J., Park Thomas, I., Schweder, P., Scotter, J., Correia, J., Smith, A., Gibbons, H., Oldfield, R., Bergin, P., Mee, E., Faull, R., Curtis, M., Scott, G. and Dragunow, M.: Isolation of Highly Enriched Primary Human Microglia for Functional Studies., Sci Rep 6, 19371, 2016 |
| 人 | Human, 25-81 yr | 脑肿瘤 / Brain tumor | Neutral Protease: 0.11 u/ml；
Collagenase Type 4: 0.05%； Hyaluronidase: 1,000 u/ml； Deoxyribonuclease I: 5 u/ml； |
HBSS | Volovitz, I, Shapira, N., Ezer, H., Gafni, A., Lustgarten, M., Alter, T., Ben-Horin, I., Barzilai, O., Shahar, T., Kanner, A., Fried, I., Veshchev, I., Grossman, R. and Ram, Z,: A Non-Aggressive, Highly Efficient, Enzymatic Method for Dissociation of Human Brain-Tumors and Brain-Tissues to Viable Single-Cells., BMC Neurosci 17, 30, 2016 |
| 人 | Human | 淀粉样蛋白原纤维 / Amyloid Fibrils | Collagenase Type 3: 0.02%； Deoxyribonuclease I: 0.001% |
50mM Tris | Scherpelz, K., Lu, J.,Tycko, R. and Meredith, S.: Preparation of Amyloid Fibrils Seeded from Brain and Meninges, Methods Mol Biol 1345, 299, 2016 |
| 人 | 脑肿瘤 / Brain tumor | Collagenase Type 1: 0.04%；
Hyaluronidase: 0.01%； Deoxyribonuclease I: 0.02%； Neutral Protease: 0.008%； |
DMEM/F12 | Hussein, D., Punjaruk, W., Storer, L., Shaw, L., Othman, R., Ottoman, R., Peet, A., Miller, S., Bandopadhyay, G., Heath, R., Kumari, R., Bowman, K., Braker, P., Rahman, R., Jones, G., Watson, S. and Lowe, J.: Pediatric Brain Tumor Cancer Stem Cells: Cell Cycle Dynamics, DNA Repair, and Etoposide Extrusion., Neuro Oncol 13, 70-83, 2011 | |
| 人 | Human | 肿瘤 / Tumor | Collagenase Type 4: 0.1%； Hyaluronidase: 0.07%； Deoxyribonuclease I: 0.04% |
see reference | Sauvageot, C., Weatherbee, J., Kesari, S., Winters, S., Barnes, J., Dellagatta, J., Ramakrishna, N., Stiles, C., Kung, A., Kieran, M. and Wen, P.: Efficacy of the HSP90 Inhibitor 17-AAG in Human Glioma Cell Lines and Tumorigenic Glioma Stem Cells.,Neuro Oncol Vol. 11,, , 109, 2009 |
| 人 | Human | 小胶质细胞 / Microglia | Trypsin: 0.25%； Deoxyribonuclease I: .005% |
DMEM/F12 | Klegeris Andis, McGeer PatrickL: Chymotrypsin-like proteases contribute to human monocytic THP-1 cell as well as human microglial neurotoxicity, Glia 51, 56-64, 2005 |
| 人 | Human | 微血管 / Microvessels | Collagenase Type 4: 0.1% | DMEM | Gerhart, D. Z., Broderius, M. A., and Drewes, L. R.: Cutlured Human and Canine Endothelial Cells from Brain Microvessels, Brain Res Bull 21, 785, 1988 |
| 人 | Human | 神经元 / Neuronal | Deoxyribonuclease I: 10 痢/ml | Tris-HCl, 50 mM, CaCl2, 2 mM | Roher, A.E., Palmer, K.C., Chau, V., and Ball, M.J.: Isolation and Chemical Characterization of Alzheimer's Disease Paired Helical Filament Cytoskeletons:Differentiation from Amyloid Plaque Core Protein, J Cell Biol 107, 2703, 1988 |
| 人 | Human, 15-54 years | 微血管 / Microvessels | Collagenase: 0.1% | Serum-free modified Lewis medium | Vinters, H.V., Reave, S., Costello, P., Girvin, J.P., and Moore, S.A.: Isolation and Culture of Cells Derived From Human Cerebral Microvessels, Cell Tissue Res 249, 657, 1987 |
| 昆虫 | Drosophilia | 神经元 / Neurons | Papain: 50 u/ml | Modified Dissecting Saline | Hadzic, T., Park, D., Abruzzi, K., Yang, L., Trigg, J., Rohs, R., Rosbash, M. and Taghert, P.: Genome-Wide Features of Neuroendocrine Regulation in Drosophila by the Basic Helix-Loop-Helix Transcription Factor DIMMED., Nucleic Acids Res 43, 2199-215, 2015 |
| 昆虫 | Drosophila | 神经元 / Neurons | Papain: 20 u/ml | Saline | Gu, H. and O'Dowd, D.: Whole Cell Recordings from Brain of Adult Drosophila., J Vis Exp 6, 248, 2007 |
| 昆虫 | Drosophila | 神经元 / Neurons | Papain: 4 u/ml | DMEM | Sicaeros, B. Campusano, J. and O'Dowd, D.: Primary Neuronal Cultures from the Brains of Late Stage Drosophila Pupae., J Vis Exp 4, 200, 2007 |
| 昆虫 | Drosophilia | 肯扬 / Kenyon | Papain: 20 u/ml | Recording saline | Gu, H. and O'Dowd, D.: Cholinergic Synaptic Transmission in Adult Drosophila Kenyon Cells in Situ., J Neurosci 26, 265-72, 2006 |
| 猴 | Monkey, rhesus | 脑细胞 / Brain cells | Collagenase Type 2: 500 u/ml； Deoxyribonuclease I: 28 u/ml |
HBSS | Marcondes MC, Burudi EM, Huitron-Resendiz S, Sanchez-Alavez M, Watry D, Zandonatti M, Henriksen SJ, Fox HS: Highly activated CD8(+) T cells in the brain correlate with early central nervous system dysfunction in simian immunodeficiency virus infection, J Immunol 167, 5429-38, 2001 |
| 小鼠 | Adult and neonatal | 小胶质细胞，星形胶质细胞，淋巴细胞 / Microglia, astrocytes, lymphocytes | Collagenase Type 2: 120 u/ml； Papain: 20 u/ml； Neutral Protease: 1.2 u/ml； Deoxyribonuclease I: 20 u/ml |
EBSS | Calvo, Belen, Rubio, F., Fernandez, M. and Tranque, P.: Dissociation of Neonatal and Adult Mice Brain for Simultaneous Analysis of Microglia, Astrocytes and Infiltrating Lymphocytes by Flow Cytometry.,IBRO Rep Vol. 8,, , 36-47, 2020 |
| 小鼠 | Mouse, adult | 星形胶质细胞，小胶质细胞，内皮细胞，少突胶质细胞 / Astrocytes, microglia, endothelia, oligodendrocytes | Papain: 20 u/ml； Collagenase Type 1: 0.01% |
HBSS | Swartzlander, D., Propson, N., Roy, E., Saito, T., Saido, T., Wang, B. and Zheng, H.: Concurrent Cell Type-Specific Isolation and Profiling of Mouse Brains in Inflammation and Alzheimer's Disease., JCI Insight 3, , 2018 |
| 小鼠 | Mouse | 神经元 / Neurons | Papain: 10 u/ml； Deoxyribonuclease I: 10 u/ml |
Artificial CSF | Kramer, D., Risso, D., Kosillo, ., Ngai, J. and Bateup, H.: Combinatorial Expression of Grp and Neurod6 Defines Dopamine Neuron Populations with Distinct Projection Patterns and Disease Vulnerability,eNeuro Vol. 5,, , 1, 2018 |
| 小鼠 | Mouse, fetal and mature | 星形胶质细胞和神经元 / Astrocytes and neurons | Papain: 7.5-20 u/ml | RPMI | Zhang, Y., Sloan, S., Clarke, L., Caneda, C., Plaza, C., Blumenthal, P., Vogel, H., Steinberg, G., Edwards, M., Li, G., Duncan, J., Cheshier, S., Shuer, L., Chang, E., Grant, G., Gephart, M. and Barres, B.: Purification and Characterization of Progenitor and Mature Human Astrocytes Reveals Transcriptional and Functional Differences with Mouse., Neuron 89, 37-53, 2016 |
| 小鼠 | Mouse | 脉络丛 / Chloroid plexus | Collagenase Type 4: 400 u/ml | PBS | Baruch, K., Rosenzweig, N., Kertser, A., Deczkowska, A., Sharif, A., Spinrad, A., Tsitsou-Kampeli, A., Sarel, A., Cahalon, L. and Schwartz, M.: Breaking Immune Tolerance by Targeting Foxp3(+) Regulatory T Cells Mitigates Alzheimer's Disease Pathology., Nat Commun 6, 7967, 2015 |
| 小鼠 | Mouse, 3 week | 神经 / Neural | Papain: 20 u/ml | PBS | Luo,Y.,Coskun, V., Liang, A.,Yu, J., Cheng, L.,Ge, W., Shi, Z., Zhang, K., Li, C., Lin, H., Luo, D., Wang, J., Lin, C., Dai, Z., Zhu, H., Zhang, J., Liu, J., Liu, H.,deVellis, J., Horvath, S., Sun, Y. and Li, S.: Different Patterns of Molecular Forms of Somatostatin are Released by the Rat Median Eminence and Hypothalamus., Cell 161, 1175, 2015 |
| 小鼠 | Mouse | 脑隔离白细胞 / Brain-sequestered leukocytes | Collagenase Type 4: 0.05%； Deoxyribonuclease I: 2 u/ml |
RPMI | Ryg-Cornejo, V., Ioannidis, L. and Hansen, D.: Isolation and Analysis of Brain-Sequestered Leukocytes from Plasmodium Berghei ANKA-Infected Mice., J Vis Exp 71, e50112, 2013 |
| 小鼠 | Mouse | 脑周细胞 / Cerebral pericytes | Collagenase Type 2: 0.1%； Deoxyribonuclease I: 30 u/ml |
DMEM | Shah. G., Price, T., Banks, W., Morofuji, Y., Kovac, A., Ercal, N., Sorenson, C., Shin, E., Sheibani, N.: Pharmacological Inhibition of Mitochondrial Carbonic Anhydrases Protects Mouse Cerebral Pericytes from High Glucose-Induced Oxidative Stress and Apoptosis., J Pharmacol Exp Ther 344, 637-45, 2013 |
| 小鼠 | Mouse, P13-15 | 胶质细胞 / Glial | Papain: 20 u/ml； Deoxyribonuclease I: 0.0005% |
EBSS | Haseleu, J., Anlauf, E., Blaess, S., Endl, E. and Derouiche, A.: Studying Subcellular Detail in Fixed Astrocytes: Dissociation of Morphologically Intact Glial Cells (DIMIGs)., Front Cell Neurosci 7, 54, 2013 |
| 小鼠 | Mouse, P0-P15 | 星形胶质细胞 / Astrocytes | Papain: 5 u/ml； Deoxyribonuclease I: 0.004% |
DPBS | Foo, L.: Purification of Rat and Mouse Astrocytes by Immunopanning., Cold Spring Harb. Protoc. 2013, 421-32, 2013 |
| 小鼠 | Mouse, adult | 神经元 / Neurons | PDS kit: with modifications | DMEM/F12 | Saxena, A., Wagatsuma, A., Noro, Y., Kuji, T., Asaka-Oba, A., Watahiki, A., Gurnot, C., Fagiolini, M., Hensch, T. and Carninci, P.: Trehalose-Enhanced Isolation of Neuronal Sub-Types from Adult Mouse Brain., Biotechniques 52, 381, 2012 |
| 小鼠 | Mouse, 1-2 day | 少突胶质细胞，背根神经节 / Oligodendrocytes, dorsal root ganglia | Papain: 0.15%； Deoxyribonuclease I: 0.006% |
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 |
| 小鼠 | Mouse, 6-12 week | 血管平滑肌细胞 / Vascular smooth muscle cells | Papain: 0.05%； Collagenase Type 4: 0.15%； Elastase: 0.05% |
PBS | Chung, W., Farley, J., Swenson, A., Barnard, J., Hamilton, G., Chiposi, R. and Drummond, H.: Extracellular Acidosis Activates ASIC-like Channels in Freshly Isolated Cerebral Artery Smooth Muscle Cells., Am J Physiol Cell Physiol 298, C1198, 2010 |
| 小鼠 | Mouse, 4-6 day | 神经元 / Neurons | PDS kit: with modifications | HBSS | Lee, H., Greene, L., Mason, C. and Manzini, M.: Isolation and Culture of Post-Natal Mouse Cerebellar Granule Neuron Progenitor Cells and Neurons., J Vis Exp 23, 990, 2009 |
| 小鼠 | Mouse, postnatal | 星形胶质细胞 / Astrocytes | Trypsin: 0.25%； Deoxyribonuclease I: 1,000 u/ml |
HBSS | Sher, F., Rossler, R., Brouwer, N., Balasubramaniyan, V., Boddeke, E. and Copray, S.: Differentiation of Neural Stem Cells Into Oligodendrocytes: Involvement of the Polycomb Group Protein Ezh2., Stem Cells 26, 2875, 2008 |
| 小鼠 | Mouse, neonatal | 神经元 / Neurons | Papain: 20 u/ml | Neurobasal | Fasano, C., Thibault, D. and Trudeau, L.: Culture of Postnatal Mesencephalic Dopamine Neurons on an Astrocyte Monolayer,Current Protocols in Neuroscience Vol. 44,, , 3.21.1, 2008 |
| 小鼠 | Mouse, embryonic and postnatal | 皮层神经元 / Cortical neurons | Papain: 4-10 u/ml | Neurobasal | Hilgenberg, L. and Smith, M.: Preparation of Dissociated Mouse Cortical Neuron Cultures., J Vis Exp 10, 562, 2007 |
| 小鼠 | Mouse | 神经球 / Neurosperes | PDS kit: per instructions | DMEM/F12 | Klein C, Butt SJ, Machold RP, Johnson JE, and Fishell G.: Cerebellum- and forebrain-derived stem cells possess intrinsic regional character, Development 132, 4497, 2005 |
| 小鼠 | Mouse, 1 day | 神经祖细胞 / Neural progenitor | PDS kit: per instructions | see reference | Seaberg, R., Smukler, S. and Van der Kooy, D.: Intrinsic Differences Distinguish Transiently Neurogenic Progenitors from Neural Stem Cells in the Early Postnatal Brain., Dev Biol 278, 71, 2005 |
| 小鼠 | Mouse | 颗粒细胞前体，前肿瘤细胞和肿瘤细胞 / Granule cell precursors, pre-neoplastic and tumor cells | Papain: 10 u/ml； Deoxyribonuclease I: 250 u/ml |
Neurobasal/B27 | Oliver, T., Read, T., Kessler, J., Mehmeti, A., Wells, J., Huynh, T., Lin, S. and Wechsler-Reya, R.: Loss of Patched and Disruption of Granule Cell Development in a Pre-Neoplastic Stage of Medulloblastoma., Development 132, 2425, 2005 |
| 小鼠 | Mouse | 皮层神经元 / Cortical neurons | PDS kit: per instructions | Neurobasal | Hernandez, F., Perez, M., Lucas, J., Mata, A., Bhat, R. and Avila, J.: Glycogen Synthase Kinase-3 plays a Crucial Role in Tau Exon 10 Splicing and Intranuclear Distribution of SC35. Implications for Alzheimer's Disease., J Biol Chem 279, 3801, 2004 |
| 小鼠 | Mouse, 3 day | 小胶质细胞 / Microglia | Papain: 90 u/ml； Deoxyribonuclease I: 2000 u/ml |
Eagle's MEM | Nishioku T, Hashimoto K, Yamashita K, Liou SY, Kagamiishi Y, Maegawa H, Katsube N, Peters C, von Figura K, Saftig P, Katunuma N, Yamamoto K, Nakanishi H: Involvement of cathepsin E in exogenous antigen processing in primary cultured murine microglia, J Biol Chem 277, 4816, 2002 |
| 小鼠 | Mouse, SD, 8-12 week | 小胶质细胞 / Microglia | Trypsin: 0.125%； Collagenase Type 2: 0.01%； Deoxyribonuclease I: .005% |
RPMI-1640 | O'Donnell SL, Frederick TJ, Krady JK, Vannucci SJ, Wood TL: IGF-I and microglia/macrophage proliferation in the ischemic mouse brain, Glia 39, 85, 2002 |
| 小鼠 | Mouse, 2-5 day | 后黑质 / Postnal substantia nigra | PDS kit: per instructions | see reference | Smeyne Michelle, Smeyne RichardJ: Method for culturing postnatal substantia nigra as an in vitro model of experimental Parkinson's disease, Brain Res Brain Res Protoc 9, 105-11, 2002 |
| 小鼠 | Mouse, embryonic | 皮质祖细胞 / Cortical progenitors | PDS kit: per instructions | Serum free medium | Estivill-Torrus, G., Pearson, H., Van Heyningen, V., Price, D. and Rashbass, P.: Pax6 is Required to Regulate the Cell Cycle and the Rate of Progression from Symmetrical to Asymmetrical Division in Mammalian Cortical Progenitors., Development 129, 455, 2002 |
| 小鼠 | Mouse, 1 day | 神经元和神经胶质 / Neurons and glia | PDS kit: per instructions | Neurobasal | Martin-Aparicio, E., Yamamoto, A., Hernandez, F., Hen, R., Avila, J. and Lucas, J.: Proteasomal-Dependent Aggregate Reversal and Absence of Cell Death in a Conditional Mouse Model of Huntington's Disease., J Neurosci 21, 8772, 2001 |
| 小鼠 | Mouse | 小脑颗粒细胞前体 / Cerebellar granule cell precursors | Papain: 0.435%； Deoxyribonuclease I: 0.05% |
EBSS | Miyazawa K, Himi T, Garcia V, Yamagishi H, Sato S, and Ishizaki Y.: A role for p27/Kip1 in the control of cerebellar granule cell precursor proliferation, J Neurosci 20, 5756, 2000 |
| 小鼠 | Mouse | 星形胶质细胞 / Astrocytes | Trypsin: 0.25% | DMEM | Lim, D, and Alvarez-Buylla, A.: Interaction Between Astrocytes and Adult Subventricular Zone Precursors Stimulates Neurogenesis, Proc Natl Acad Sci U S A 96, 7526, 1999 |
| 小鼠 | Mouse, newborn | 海马细胞 / Hippocampal cells | Papain: 10 u/ml | DMKM | Jun K, Choi G, Yang SG, Choi KY, Kim H, Chan GC, Storm DR, Albert C, Mayr GW, Lee CJ, Shin HS: Enhanced hippocampal CA1 LTP but normal spatial learning in inositol 1,4,5-trisphosphate 3-kinase(A)-deficient mice, Learn Mem 5, 317-30, 1998 |
| 小鼠 | Mouse, SWR or CF1, 1-3 months | 乳头，味觉感受器 / 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 |
| 小鼠 | Mouse, 0-30 day | 神经 / Neural | Trypsin NF 1:250: 0.25% | BSS | Shrier, B., Wilson, S., and Nirenberg, M.: Cultured Cell Systems and Methods for Neurobiology, Vol. 32,, , 765, 1974 |
| 绵羊 | Ovine | 神经元 / Neurons | Papain: 0.2%； Deoxyribonuclease I: 0.1% |
Hibernate | Reddy, R., Amodei, R., Estill, C., Stormshak, F., Meaker, M. and Roselli, C.: Effect of Testosterone on Neuronal Morphology and Neuritic Growth of Fetal Lamb Hypothalamus-Preoptic Area and Cerebral Cortex in Primary Culture., PLoS ONE 10, e0129521, 2015 |
| 绵羊 | Ovine | 神经元 / Neurons | PDS kit: per instructions | DMEM | Lepore G, Gadau S, Mura A, Zedda M, Farina V: Aromatase Immunoreactivity in Fetal Ovine Neuronal Cell Cultures Exposed to Oxidative Injury., Eur J Histochem 53, e28, 2009 |
| 绵羊 | Lamb | 少突胶质细胞，神经 / Oligodendroglia，Neural | 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 |
| 猪 | Mini pigs, Yucatan (Susscrofa Yucatan), 4-6 months | 微血管 / Microvascular | Collagenase: 0.1% | HBSS | Robinson, D.H., Kang,Y., Deschner, S.H., and Nielsen, T.B.: Morphologic Plasticity and Periodicity : Porcine Cerebral Microvascular Cells in Culture, In Vitro Cell Dev Biol 26, 169, 1990 |
| 大鼠 | Rat, SD, 1-2 day | 少突胶质前体细胞 / Oligodendrocyte precursor cells | Papain: 16.5 u/ml | HBSS | Kurachi, M., Mikuni, M. and Ishizaki, Y.: Extracellular Vesicles from Vascular Endothelial Cells Promote Survival, Proliferation and Motility of Oligodendrocyte Precursor Cells., PLoS ONE 11, e0159158, 2016 |
| 大鼠 | Rat, SD, E18 | 神经元 / Neurons | Papain: 20 u/ml | HBSS | Folts, C., Scott-Hewitt, N., Proschel, C., Mayer-Proschel, M. and Noble, M.: Lysosomal Re-acidification Prevents Lysosphingolipid-Induced Lysosomal Impairment and Cellular Toxicity., PLoS Biol 14, e1002583, 2016 |
| 大鼠 | Rat, P0-P15 | 星形胶质细胞 / Astrocytes | Papain: 5 u/ml； Deoxyribonuclease I: 0.004% |
DPBS | Foo, L.: Purification of Rat and Mouse Astrocytes by Immunopanning., Cold Spring Harb. Protoc. 2013, 421-32, 2013 |
| 大鼠 | Rat, SD, neonatal | 神经元 / Neurons | Papain: 20 u/ml | Neuro medium | Kaiser, O., Aliuos, P., Wissel, K., Lenarz, T., Werner, D., Reuter, G., Kral, A. and Warnecke, A.: Dissociated Neurons and Glial Cells Derived from Rat Inferior Colliculi After Digestion with Papain., PLoS ONE 8, e80490, 2013 |
| 大鼠 | Rat, SD, 19-21 day | 视交叉上核神经元 / Suprachiasmatic nucleus neurons | Papain: 100 u/ml | MEM | Cao, R., Li, A., Cho, H., Lee, B. and Obrietan, K.: Mammalian Target of Rapamycin Signaling Modulates Photic Entrainment of the Suprachiasmatic Circadian Clock., J Neurosci 30, 6302, 2010 |
| 大鼠 | Rat, Fisher, 7-21 month | 海马神经元 / 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 | Rat, SD, 7 day | 小脑颗粒神经元 / 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 | Rat, neonatal | 星形胶质细胞 / Astrocytes | Papain: 20 u/ml | EBSS | Shigetomi, E. and Khakh, B.: Measuring Near Plasma Membrane and Global Intracellular Calcium Dynamics in Astrocytes., J Vis Exp 26, 1142, 2009 |
| 大鼠 | Rat, neonatal | 海马神经元 / Hippocampal neurons | Papain: 10 u/ml | EBSS | Richler Esther, Chaumont Severine, Shigetomi Eiji, Sagasti Alvaro, Khakh Baljit S: Tracking transmitter-gated P2X cation channel activation in vitro and in vivo, Nat Methods 5, 87-93, 2008 |
| 大鼠 | Rat, SD | 海马 / Hippocampal | Trypsin: 0.1% | Neurobasal | Akanda, N., Tofighi, R., Brask, J., Tamm, C., Elinder, F. and Ceccatelli, S.: Voltage-Dependent Anion Channels (VDAC) in the Plasma Membrane Play a Critical Role in Apoptosis in Differentiated Hippocampal Neurons but not in Neural Stem Cells., Cell Cycle 7, 3225, 2008 |
| 大鼠 | Rat, neonatal | 硬脑膜 / Dura mater | Collagenase Type 1: 0.075% | DMEM | Peptan, I., Hong, L. and Evans, C.: Multiple Differentiation Potentials of Neonatal Dura Mater-Derived Cells., Neurosurgery 60, 346, 2007 |
| 大鼠 | Rat, E18 | 海马神经元 / Hippocampal neurons | Papain: 0.2% | Hibernate | Jekabsons MB, Nicholls DG: Bioenergetic analysis of cerebellar granule neurons undergoing apoptosis by potassium/serum deprivation, Cell Death Differ 13, 1595-610, 2006 |
| 大鼠 | Rat, Wistar, 1-3 day | 海马神经元 / Hippocampal neurons | Trypsin: 0.05% | DMEM | Velasco Myrian, Garci苔 Esperanza, Onetti Carlos G: Glucose deprivation activates diversity of potassium channels in cultured rat hippocampal neurons, Cell Mol Neurobiol 26, 307-19, 2006 |
| 大鼠 | Rat, E19 | 海马神经元 / Hippocampal neuron | Papain: 10 u/ml | MEM | Khakh Baljit S, Fisher James A, Nashmi Raad, Bowser David N, Lester Henry A: An angstrom scale interaction between plasma membrane ATP-gated P2X2 and alpha4beta2 nicotinic channels measured with fluorescence resonance energy transfer and total internal reflection fluorescence microscopy, J Neurosci 25, 6911-20, 2005 |
| 大鼠 | Rat, Wistar, 14 day | 视觉皮层 / Visual cortical | PDS kit: per instructions | EBSS | Mizoguchi Y, Kanematsu T, Hirata M, Nabekura J: A rapid increase in the total number of cell surface functional GABAA receptors induced by brain-derived neurotrophic factor in rat visual cortex, J Biol Chem 278, 44097, 2003 |
| 大鼠 | Rat, Wistar, male | 脑内皮 / Cerebral endothelial | Collagenase Type 3: 0.2% | MEM | Floris S, Van den Born J, van der Pol SM, Dijkstra CD, De Vries HE: Heparan sulfate proteoglycans modulate monocyte migration across cerebral endothelium, J Neuropathol Exp Neurol 62, 780, 2003 |
| 大鼠 | Rat, SD, E18 | 皮质细胞 / Cortical cells | Papain: 20 u/ml； Deoxyribonuclease I: .005% |
EBSS | Behar, T, Smith, S, Kennedy, R, Mckenzie, J, Maric, I and Barker, J: GABA(B) receptors mediate motility signals for migrating embryonic cortical cells, Cereb Cortex 11, 744-53, 2001 |
| 大鼠 | Rat, wistar, 250-300 g | 脑动脉平滑肌细胞 / Cerebral artery smooth muscle cells | Papain: 1.5 mg/ml； Collagenase Type 4: 1.5 mg/ml |
Physiological Salt Solution | Brzezinska AK, Gebremedhin D, Chilian WM, Kalyanaraman B, Elliott SJ.: Peroxynitrite reversibly inhibits Ca(2+)-activated K(+) channels in rat cerebral artery smooth muscle cells, Am J Physiol Heart Circ Physiol 278, H1883, 2000 |
| 大鼠 | Rat, SD, E16 | 皮层神经元 / Cortical neurons | PDS kit: with modifications | Neurobasal Medium | Varney MA, Cosford ND, Jachec C, Rao SP, Sacaan A, Lin FF, Bleicher L, Santori EM, Flor PJ, Allgeier H, Gasparini F, Kuhn R, Hess SD, Velicelebi G, Johnson EC: SIB-1757 and SIB-1893: selective, noncompetitive antagonists of metabotropic glutamate receptor type 5, J Pharmacol Exp Ther 290, 170, 1999 |
| 大鼠 | Rat, wistar, neonatal | 海马细胞 / Hippocampal cells | Papain: 1 mg/ml | DMEM | Boehm S, Betz H: Somatostatin inhibits excitatory transmission at rat hippocampal synapses via presynaptic receptors, J Neurosci 17, 4066, 1997 |
| 大鼠 | Rat, SD, Fisher | 海马神经元 / 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 | Rat, E18 | 海马和皮层神经元 / Hippocampal and cortical neurons | PDS kit: per instructions | Neurobasal media | Naeve GS, Ramakrishnan M, Kramer R, Hevroni D, Citri Y, Theill LE: Neuritin: a gene induced by neural activity and neurotrophins that promotes neuritogenesis, Proc Natl Acad Sci U S A 94, 2648-53, 1997 |
| 大鼠 | Rat, SD, newborn | 胶质细胞 / Glial | Trypsin: 0.0625% | MEM, sterile | Pixley, S.K.: The Olfactory Nerve Contains Two Populations of Glia, In Vitro 5, 269, 1992 |
| 大鼠 | Rat, newborn | 星形胶质细胞 / Astrocytes | Trypsin: 0.25% | DMEM, HBSS | Holzwarth, J., Glaum, S., and Miller, R.: Activation of Endothelin Receptors by Sarafotoxin Regulates Ca2+ Homeostasis in Cerebellar Astrocytes, Glia 5, 239, 1992 |
| 大鼠 | Rat, 60-72 hours old | 成纤维细胞 / Fibroblasts | Trypsin: 0.2% | HEPES buffered DMEM | Acheson, A., Barker, P., Alderson, R., Miller, F., and Murphy, R.: Detection of Brain-Derived Neurotrophic Factor-like Activity in Fibroblasts and Schwann Cells: Inhibition by Antibodies to NGF, Neuron 7, 265, 1991 |
| 大鼠 | Rat, SD, adult | 内皮 / Endothelial | Collagenase Type 2: 0.5% | Medium 199 | Doron, D., Jacobowitz, D., Heldman, E., Feurerstein, G., Pollard, H., and Hallenbeck, J.: Extracellular Matrix Permits the Expression of Von Willebrand's Factor, Uptake of Di-I-Acetylated Low Density Lipoprotein and Secretion of Prostacyclin in Cultures of Endothelial Cells from Rat Brain Microvessels, In Vitro Cell Dev Biol 27A, 689, 1991 |
| 大鼠 | Rat, SD, 5 day | 海马神经元 / Hippocampal neurons | Trypsin: 0.2%； Deoxyribonuclease I: 10 ug/ml |
HBSS | Mattson, M., and Kater, S.: Development and Selective Neurodegeneration in Cell Cultures From Different Hippocampal Regions, Brain Res 490, 110, 1989 |
| 大鼠 | Rat, SD, female | 胎鼠大脑 / Fetal rat brain | Collagenase Type 4: 0.1% | DMEM | Matsuda, M.: Serum Proteins Enhance Aggregate Formation of Dissociated Fetal Rat Brain Cells in an Aggregating Culture, In Vitro Cell Dev Biol 24 (10), 1031, 1988 |
| 大鼠 | Rat, albino, adult and newborn | 大脑皮质 / Cerebral cortices | Trypsin: 0.25% | PBS | Giulian, D. and Baker, T.J.: Characterization of Ameboid Microglia Isolated From Developing Mammalian Brain, J Neurosci 6, 2163, 1986 |
| 大鼠 | Rat, Wistar, newborn | 生发基质细胞 / Germinal matrix | Trypsin: 0.25% | HBSS | Goldman, J.E., Geier, S.S., and Hirano, M.: Differentiation of Astrocytes and Oligodendrocytes From Germinal Matrix Cells in Primary Culture, J Neurosci 6, 52, 1986 |
| 大鼠 | Rat, fetus, 18-20 day | 海马神经元 / 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 | Rat, fetus, 18 day | 海马 / Hippocampi | Trypsin: 0.1% | HBSS | Rothman, S.: Synaptic Release of Excitatory Amino Acid Neurotransmitter Mediates Anoxic Neuronal Death, J Neurosci 4 (7), 1884, 1984 |
| 大鼠 | Rat, SD, 19-20 days pregnant | 神经 / Neural | Trypsin: 0.25% | DMEM | Ahmed, Z., Walker, P., and Fellows, R.: Properties of Neurons from Dissociated Fetal Rat Brain in Serum-Free Culture, J Neurosci 3 (12), 2448, 1983 |
| 大鼠 | Rat, SD, fetal | 大脑皮层，下丘脑 / Cerebral cortex，Hypothalmus | Deoxyribonuclease I: 0.001% | HEPES | Peterfreund, R. and Vale, W.: High Molecular Weight Somatostatin Secretion by Cultured Rat Brain Cell, Brain Res 239, 463, 1982 |
| 大鼠 | Rat, Wistar-Kyoto, male, 100 - 200 g | 内皮，脑 / Endothelial，Cerebral | Collagenase Type 2: 0.05% | HBSS | Diglio, C.A., Grammas, P., Filiberto Giacomelli, M.S., and Wiener, J.: Primary Culture of Rat Cerebral Microvascular Endothelial Cells, Lab Invest 46, 554, 1982 |
| 大鼠 | Rat, fetus | 皮质 / Cortical | Trypsin: 0.25% | Puck's D1 (see reference) | Swaiman, K., Neale, E., Fitzgerald, S., and Nelson, P.: A Method for Large-scale Production of Mouse Brain Cortical Cultures, Brain Res 255, 361, 1982 |
| 大鼠 | Rat, Wistar, pregnant | 胶质细胞 / Glial | Trypsin: 0.05% | Eagle's MEM/DMEM | Abney, E., Bartlett, P., and Raff, M.: Astrocytes, Ependymal Cells, and Oligodendrocytes Develop on Schedule in Dissociated Cell Cultures of Embryonic Rat Brain, Dev Biol 83, 301, 1981 |
| 大鼠 | Rat, SD, one month old | 毛细血管内皮，周细胞 / Capillary endothelium，Pericytes | Neutral Protease: 0.1% | Medium 199 | Bowman, P., Betz, A., Ar, D., Wolinsky, J., Penney, J., Shivers, R., and Goldstein, G.: Primary Culture of Capillary Endothelium From Rat Brain, In Vitro 17 (4), 353, 1981 |
| 大鼠 | Rat, Wistar, male, 300-500 g | 微血管，内皮 / Microvessels，Endothelial | Collagenase Type 2: 0.75% | Ringers-HEPES buffer | Williams, S., Gillis, J., Matthews, M., Wagner, R., and Bitensky, M.: Isolation and Characterization of Brain Endothelial Cells: Morphology and Enzyme Activity, J Neurochem 35 (2), 374, 1980 |
| 大鼠 | Rat, Wistar, adult, 170 g | 内皮 / Endothelial | Trypsin: 0.5% | BSS | Phillips, P., Kumar, P., Kumar, S., and Waghe, M.: Isolation And Characterization of Endothelial Cells From Rat And Cow Brain White Matter, J Anat 129, 261, 1979 |
| 大鼠 | Rat, Holtzmann, 18 day | 海马神经元 / Hippocampal neurons | Trypsin: 0.1% | HBSS | Banker, G., and Cowan, M.: Rat Hippocampal Neurons in Dispersed Cell Culture, Brain Res 126, 397, 1977 |
| 大鼠 | Rat, SD, 2 days | 松果体 / Pineal | Trypsin: 0.25% | DMEM, MEM | Rowe, V., Neale, E., Avins, L., Guroff, G., and Schrier, B.: Pineal gland cells in culture. Morphology, Biochemistry, Differentiation, and co-culture with sympathetic neurons, Exp Cell Res 104, 345, 1977 |
| 贝类 | Helisoma trivolvis | 神经元，颊神经节 / Neurons，Buccal ganglia | Trypsin: 0.2% | Saline, Sterile | Hadley, R.D., Bodnar, D.A., and Kater, S.B.: Formation of Electrical Synapses Between Isolated, Cultured Helisoma Neurons Requires Mutual Neurite Elongation, J Neurosci 5, 3145, 1985 |
