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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

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
Bovine微血管内皮 / Microvascular endothelialNeutral Protease: 0.005%Medium 199Kanda, 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
Bovine脑内皮细胞 / Brain endothelial cellsCollagenase Type 2: 0.35%DMEMWolburg 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
Bovine内皮 / EndothelialNeutral Protease: 0.125%MEMMiller, 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
Bovine, adult脑动脉,内皮 / Cerebral artery,EndothelialCollagenase: 0.2%HBSSMachi, 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
Bovine毛细血管内皮 / Capillary endothelialCollagenase: 0.1%DMEMEstrada, C., Bready, J., Berliner, J., and Cancilla, P.: Choline Uptake by Cerebral Capillary Endothelial Cells in Culture, J Neurochem 54, 1467, 1990
Bovine内皮 / EndothelialNeutral Protease: 0.5%MEMAudus, 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
Bovine内皮,脑动脉 / Endothelial,Brain arteriesCollagenase Type 2: 0.2%Dulbecco's PBSGoetz, 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
Calf少突胶质细胞,神经 / Oligodendroglia,NeuralTrypsin: 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 / NeuronsTrypsin: 0.06-0.08%PIPES salineKay, 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
Human小胶质细胞 / MicrogliaCollagenase Type 1: 300 u/ml;
Trypsin: 0.125%
DMEMMizee, 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
Human, adult神经元 / NeuronalPapain: 20 u/mlNeurobasalSpaethling, 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
Human, fetal and mature星形胶质细胞和神经元 / Astrocytes and neuronsPapain: 7.5-20 u/mlRPMIZhang, 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
Human小胶质细胞 / MicrogliaNeutral Protease: 10 u/ml;
Papain: 2.5 u/ml
Hibernate ARustenhoven, 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
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;

HBSSVolovitz, 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
Human淀粉样蛋白原纤维 / Amyloid FibrilsCollagenase Type 3: 0.02%;
Deoxyribonuclease I: 0.001%
50mM TrisScherpelz, K., Lu, J.,Tycko, R. and Meredith, S.: Preparation of Amyloid Fibrils Seeded from Brain and Meninges, Methods Mol Biol 1345, 299, 2016

脑肿瘤 / Brain tumor

Collagenase Type 1: 0.04%;

Hyaluronidase: 0.01%;

Deoxyribonuclease I: 0.02%;

Neutral Protease: 0.008%;

DMEM/F12Hussein, 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
Human肿瘤 / TumorCollagenase Type 4: 0.1%;
Hyaluronidase: 0.07%;
Deoxyribonuclease I: 0.04%
see referenceSauvageot, 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
Human小胶质细胞 / MicrogliaTrypsin: 0.25%;
Deoxyribonuclease I: .005%
DMEM/F12Klegeris Andis, McGeer PatrickL: Chymotrypsin-like proteases contribute to human monocytic THP-1 cell as well as human microglial neurotoxicity, Glia 51, 56-64, 2005
Human微血管 / MicrovesselsCollagenase Type 4: 0.1%DMEMGerhart, D. Z., Broderius, M. A., and Drewes, L. R.: Cutlured Human and Canine Endothelial Cells from Brain Microvessels, Brain Res Bull 21, 785, 1988
Human神经元 / NeuronalDeoxyribonuclease I: 10 痢/mlTris-HCl, 50 mM, CaCl2, 2 mMRoher, 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
Human, 15-54 years微血管 / MicrovesselsCollagenase: 0.1%Serum-free modified Lewis mediumVinters, 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 / NeuronsPapain: 50 u/mlModified Dissecting SalineHadzic, 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 / NeuronsPapain: 20 u/mlSalineGu, 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 / NeuronsPapain: 4 u/mlDMEMSicaeros, 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 / KenyonPapain: 20 u/mlRecording salineGu, H. and O'Dowd, D.: Cholinergic Synaptic Transmission in Adult Drosophila Kenyon Cells in Situ., J Neurosci 26, 265-72, 2006
Monkey, rhesus脑细胞 / Brain cellsCollagenase Type 2: 500 u/ml;
Deoxyribonuclease I: 28 u/ml
HBSSMarcondes 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, lymphocytesCollagenase Type 2: 120 u/ml;
Papain: 20 u/ml;
Neutral Protease: 1.2 u/ml;
Deoxyribonuclease I: 20 u/ml
EBSSCalvo, 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, oligodendrocytesPapain: 20 u/ml;
Collagenase Type 1: 0.01%
HBSSSwartzlander, 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 / NeuronsPapain: 10 u/ml;
Deoxyribonuclease I: 10 u/ml
Artificial CSFKramer, 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 neuronsPapain: 7.5-20 u/mlRPMIZhang, 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 plexusCollagenase Type 4: 400 u/mlPBSBaruch, 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


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