worthington 脑组织分离指南

发布者：艾美捷科技发布时间：2022-05-31

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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：脑组织分离指南

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。

脑
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