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worthington 神经系统组织分离指南

发布者:艾美捷科技    发布时间:2022-06-07     
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原代细胞分离中,针对神经系统组织涉及到中肠DRG,睫状神经节神经元、心脏,肾上腺嗜铬,副神经元、卫星胶质等等的解离通常会使用酶,如何选择合适的酶来完成对应神经系统组织的解离,以及原代细胞采集的常规操作, 艾美捷针对神经系统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:worthington神经系统组织分离指南


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

神经系统
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
禽类Quail and Chicken, embryonic中肠DRG / Midgut DRGCollagenase animal free: 0.05%;
Neutral Protease: 0.5%
Ham's F12Rollo, B., Zhang, D., Simkin, J., Menheniott, T. and Newgreen, D.: Why are Enteric Ganglia so Small? Role of Differential Adhesion of Enteric Neurons and Enteric Neural Crest Cells., F1000Res 4, 113, 2015
禽类Chick, White Leghorn, embryos睫状神经节神经元 / Cilary ganglion neuronsCollagenase Type 1: 0.1%PBSTemburni, 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
Bovine心脏,肾上腺嗜铬,副神经元 / Heart,Adrenal chromaffin,ParaneuronsTrypsin: 0.06%25mM HEPES buffered Locke's solution, CMFTrifaro, 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
Bovine微血管内皮 / Microvascular endothelialCollagenase/Dispase: 0.1%MEMBowman, P., Betz, A., and Goldstein, G.: Primary Culture of Microvascular Endothelial Cells From Bovine Retina, In Vitro 18 (7), 626, 1982
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
犬/狗Canine卫星胶质 / Satellite glialHyaluronidase: 0.2%;
Collagenase Type 4: 0.2%;
Collagenase: 0.2%
HBSSTongtako, W., Lehmbecker, A., Wang, Y., Hahn, K., Baumgartner, W. and Gerhauser, I.: Canine Dorsal Root Ganglia Satellite Glial Cells Represent an Exceptional Cell Population with Astrocytic and Oligodendrocytic Properties., Sci Rep 7, 13915, 2017
Chick, White Leghorn, embryos胰岛 / IsletsTrypsin: Trypsin: 0.125-0.25%;
Collagenase: 0.2%;
Deoxyribonuclease I: 200 u/ml
Eurocollins solutionWatt, P., Mullen, Y., Benhamou, P., Hober, C., Nomura, Y., Watanabe, Y., Passaro, E., Zinner, M., and Brunicardi, F.: Simplified Semiautomated Method For Isolating Islets From the Human Pancreas, Transplant Proc 26 (2), 582, 1994
Chicken, White Leghorn, embryos, 17-21 day神经元 / NeuronsPapain: 40 u/mlHEPESRaman, 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
Chick, embryo, 10-14 day平坦,视网膜 / Flat, retinaTrypsin: 0.1%Tyrode's solution, CMFMoyer, M., Bullrich, F., and Sheffield, J.: Emergence of Flat Cells From Glia in Stationary Cultures of Embryonic Chick Neural Retina, In Vitro Cell Dev Biol 26, 1073, 1990
Chick, White Leghorn, embryo脊髓 / Spinal cordTrypsin: 0.05%Phosphate bufferSchnaar, R., and Schaffner, A.: Separation of cell types from embryonic chicken and rat spinal cord: characterization of motoneuron-enriched fractions, J Neurosci 1, 204, 1981
Chick背根神经节神经元,脊髓 / Dorsal root ganglion neurons,Spinal cordTrypsin: 0.1%Puck's saline, CMFChoi, D., and Fiscbach, G.: GABA Conductance of Chick Spinal Cord and Dorsal Root Ganglion Neurons in Cell Culture, J Neurophysiol 45, 605, 1981
Chick, White Leghorn embryo, 8 day神经元,神经节 / Neurons, gangliaTrypsin: 0.25%HBSS, CMFBottenstein, J., Skaper, S., Varon, S., and Sato, G.: Selective Survival of Neurons from Chick Embryo Sensory Ganglionic Dissociates Utilizing Serum-Free Supplemented Medium, Exp Cell Res 125, 183, 1980
Chicken, White Leghorn, fertile eggs睫状神经节神经元 / Ciliary ganglion neuronsTrypsin: 0.25%Eagle's MEMTuttle, J., Suszkiw, J., and Ard, M.: Long-Term Survival and Development of Dissociated Parasympathetic Neurons in Culture, Brain Res 183, 161, 1980
Chicken, embryos, 9-10 days old背根神经节神经元 / Dorsal root ganglia neuronsCollagenase: 0.01%Eagle's MEMMudge, 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
Chick embryos神经节链,交感神经节 / Ganglion chains, sympathetic gangliaTrypsin: 0.25%Krebs PhosphosalineMcCarthy, K., and Partlow, L.: Preparation of Pure Neuronal and Non-Neuronal Cultures From Embryonic Chick Sympathetic Ganglia: A New Method Based on Both Differential Cell Adhesiveness and the Formation of Homotypic Neuronal Aggregates, Brain Res 114, 391, 1976
Chick, White Leghorn, embryos, 7 day神经视网膜 / Neural retina

Collagenase: 0.25%;

Elastase: 0.2%;

Hyaluronidase: 1.0%;

Papain: 1.0%;

Protease: 0.1%;

Trypsin: 0.05%;

HBSSWiseman, L., and Hammond, W.: The Reacquisition of Cell Adhesiveness Following Tissue Disaggregation by Eleven Different Agents, J Exp Zool 197, 429, 1976
Zebrafish, embryonic运动神经元 / MotorneuronsPDS kit: per instructionsL-15Spiro, Z., Koh, A., Tay, S., See, K. and Winkler, C.: Transcriptional Enhancement of Smn Levels in Motoneurons is Crucial for Proper Axon Morphology in Zebrafish., Sci Rep 6, 27470, 2016
Zebrafish运动神经元 / Motor neuronsCollagenase Type 2: 0.1%See referenceSakowski, S., Lunn, J., Busta, A., Palmer, M., Dowling, J. and Feldman, E.: A Novel Approach to Study Motor Neurons from Zebrafish Embryos and Larvae in Culture., J Neurosci Methods 205, 277-82, 2012
Zebrafish, Dania rerio, embryoRohon-Beard 神经元 / Rohon-Beard neuronsTrypsin: 0.2%L-15/HepesWon, 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
Zebrafish, Danio rerio, embryo神经元 / NeuronsPDS kit: with modificationsEBSSCerda, G., Hargrave, M. and Lewis, K.: RNA Profiling of FAC-sorted Neurons from the Developing Zebrafish Spinal Cord., Dev Dyn 238, 150-61, 2009
Black ghose knige fish, adult (Apteronotus albifrons)神经元,脊髓 / Neurons, spinal cordTrypsin: 0.4%PBS, CMFAnderson, M.J.: Differences in Growth of Neurons from Normal and Regenerated Teleost Spinal Cord in vitro, In Vitro Cell Dev Biol 29A, 145, 1993
青蛙Xenopus, embryonic神经元 / NeuronCollagenase Type 1: 0.1%Steinberg's solutionTakahashi, 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Guinea-pig, newborn神经元,肠 / Neuron, entericTrypsin: 0.125%Medium 199Jessen, K, McConnell, J., Purves, R., Burnstock, G., and Chamley-Campbell, J.: Tissue Culture of Mammalian Enteric Neurons, Brain Res 152, 573, 1978
仓鼠Hamster, male犁鼻器神经元 / Vomeronasal organ neuronsCollagenase Type 1: 0.02%;
Trypsin: 0.02%
PBSLiman ER: Regulation by voltage and adenine nucleotides of a Ca2+-activated cation channel from hamster vomeronasal sensory neurons, J Physiol 548, 777, 2003
Human脊髓神经祖细胞 / Spinal cord neural progenitorPDS kit: with modificationsEBSSMothe, A. and Tator, C.: Isolation of Neural Stem/Progenitor Cells from the Periventricular Region of the Adult Rat and Human Spinal Cord., J Vis Exp , e52732, 2015
Human神经的,各种 / Neural, variousPapain: 12 u/ml;
Trypsin: see reference;
Collagenase/Dispase: see reference
see referencePanchision 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
Human, 9 month神经元 / NeuronsCollagenase Type 4: 1.33%;
Papain: 0.07 u/ml;
Neutral Protease: 1 mg/ml
DMEM/F12Dietrich 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
Human, 26 week神经祖细胞 / Neural progenitor cellsPapain: 2.5 u/ml;
Deoxyribonuclease I: 250 u/ml;
Neutral Protease: 1 u/ml
DMEM/F-12Fuja 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
Human, adult心室上皮 / Ventricular epithelialPapain: 11.4 u/ml;
Deoxyribonuclease I: 10 u/ml
DMEM/F12Roy NS, Benraiss A, Wang S, Fraser RA, Goodman R, Couldwell WT, Nedergaard M, Kawaguchi A, Okano H, Goldman SA: Promoter-targeted selection and isolation of neural progenitor cells from the adult human ventricular zone, J Neurosci Res 59, 321, 2000
Human, 5-65 years视网膜色素上皮 (RPE) / Retinal pigment epithelial (RPE)Trypsin: 0.25%HBSSVon 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
Human背根神经节神经元 / Dorsal root ganglion neuronsCollagenase Type 1: 0.2%;
Neutral Protease: 0.5%
DMEM/F-12Dib-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
昆虫Drosophila, larval神经母细胞 / NeuroblastsCollagenase Type 1: 0.1%;
Papain: 0.1%
Rinaldini solutionBerger, 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
昆虫Drosophila树突 / DendritesCollagenase: 0.05%;
Neutral Protease: 0.2%
HBSSSanchez-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
昆虫Gryllus Bimaculatus巨大的中间神经元 / Giant interneuronsCollagenase: 0.05%;
Neutral Protease: 0.2%
Leibovitz's L15Kloppenburg, P. and Horner, M.: Voltage-Activated Currents in Identified Giant Interneurons Isolated from Adult Crickets Gryllus Bimaculatus, J Exp Biol 201 (Pt 17), 2529, 1998


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