worthington 神經(jīng)系統(tǒng)組織分離指南

發(fā)布者：艾美捷科技發(fā)布時(shí)間：2022-06-07

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原代細(xì)胞分離中，針對(duì)神經(jīng)系統(tǒng)組織涉及到中腸DRG，睫狀神經(jīng)節(jié)神經(jīng)元、心臟，腎上腺嗜鉻，副神經(jīng)元、衛(wèi)星膠質(zhì)等等的解離通常會(huì)使用酶，如何選擇合適的酶來(lái)完成對(duì)應(yīng)神經(jīng)系統(tǒng)組織的解離，以及原代細(xì)胞采集的常規(guī)操作， 艾美捷針對(duì)神經(jīng)系統(tǒng)相關(guān)的組織分類解離分種屬、不同的細(xì)胞、酶組合推薦，推薦濃度、培養(yǎng)基以及參考文獻(xiàn)，詳細(xì)匯總指南： worthington神經(jīng)系統(tǒng)組織分離指南

本技術(shù)指南描述了標(biāo)準(zhǔn)實(shí)驗(yàn)室程序；為建立細(xì)胞分離方案提供了一種合乎邏輯的實(shí)驗(yàn)方法；并列出了許多組織特異性參考資料，以幫助開(kāi)發(fā)有效的方法。更多其他組織的解離指南，點(diǎn)擊查看：各組織分離指南。

神經(jīng)系統(tǒng)
種屬	種屬詳情	細(xì)胞名稱	酶組合推薦，含推薦濃度	培養(yǎng)基	參考文獻(xiàn)
禽類	Quail and Chicken, embryonic	中腸DRG / Midgut DRG	Collagenase animal free: 0.05%； Neutral Protease: 0.5%	Ham's F12	Rollo, 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
禽類	Chick, White Leghorn, embryos	睫狀神經(jīng)節(jié)神經(jīng)元 / 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	Bovine	心臟，腎上腺嗜鉻，副神經(jīng)元 / Heart，Adrenal chromaffin，Paraneurons	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
牛	Bovine	微血管內(nèi)皮 / Microvascular endothelial	Collagenase/Dispase: 0.1%	MEM	Bowman, P., Betz, A., and Goldstein, G.: Primary Culture of Microvascular Endothelial Cells From Bovine Retina, In Vitro 18 (7), 626, 1982
牛	Calf	少突膠質(zhì)細(xì)胞，神經(jīng) / 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
犬/狗	Canine	衛(wèi)星膠質(zhì) / Satellite glial	Hyaluronidase: 0.2%； Collagenase Type 4: 0.2%； Collagenase: 0.2%	HBSS	Tongtako, 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
雞	Chick, White Leghorn, embryos	胰島 / Islets	Trypsin: Trypsin: 0.125-0.25%； Collagenase: 0.2%； Deoxyribonuclease I: 200 u/ml	Eurocollins solution	Watt, 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
雞	Chicken, White Leghorn, embryos, 17-21 day	神經(jīng)元 / 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	Chick, embryo, 10-14 day	平坦，視網(wǎng)膜 / 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 Embryonic Chick Neural Retina, In Vitro Cell Dev Biol 26, 1073, 1990
雞	Chick, White Leghorn, embryo	脊髓 / Spinal cord	Trypsin: 0.05%	Phosphate buffer	Schnaar, 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
雞	Chick	背根神經(jīng)節(jié)神經(jīng)元，脊髓 / Dorsal root ganglion neurons，Spinal cord	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
雞	Chick, White Leghorn embryo, 8 day	神經(jīng)元，神經(jīng)節(jié) / Neurons, ganglia	Trypsin: 0.25%	HBSS, CMF	Bottenstein, 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
雞	Chicken, White Leghorn, fertile eggs	睫狀神經(jīng)節(jié)神經(jīng)元 / 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	Chicken, embryos, 9-10 days old	背根神經(jīng)節(jié)神經(jīng)元 / 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	Chick embryos	神經(jīng)節(jié)鏈，交感神經(jīng)節(jié) / Ganglion chains, sympathetic ganglia	Trypsin: 0.25%	Krebs Phosphosaline	McCarthy, 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
雞	Chick, White Leghorn, embryos, 7 day	神經(jīng)視網(wǎng)膜 / Neural retina	Collagenase: 0.25%； Elastase: 0.2%； Hyaluronidase: 1.0%； Papain: 1.0%； Protease: 0.1%； Trypsin: 0.05%；	HBSS	Wiseman, L., and Hammond, W.: The Reacquisition of Cell Adhesiveness Following Tissue Disaggregation by Eleven Different Agents, J Exp Zool 197, 429, 1976
魚(yú)	Zebrafish, embryonic	運(yùn)動(dòng)神經(jīng)元 / Motorneurons	PDS kit: per instructions	L-15	Spiro, 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
魚(yú)	Zebrafish	運(yùn)動(dòng)神經(jīng)元 / 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 Embryos and Larvae in Culture., J Neurosci Methods 205, 277-82, 2012
魚(yú)	Zebrafish, Dania rerio, embryo	Rohon-Beard 神經(jīng)元 / 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(yú)	Zebrafish, Danio rerio, embryo	神經(jīng)元 / 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(yú)	Black ghose knige fish, adult (Apteronotus albifrons)	神經(jīng)元，脊髓 / Neurons, spinal cord	Trypsin: 0.4%	PBS, CMF	Anderson, 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	神經(jīng)元 / 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	Guinea-pig, newborn	神經(jīng)元，腸 / 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
倉(cāng)鼠	Hamster, male	犁鼻器神經(jīng)元 / Vomeronasal organ neurons	Collagenase Type 1: 0.02%； Trypsin: 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
人	Human	脊髓神經(jīng)祖細(xì)胞 / Spinal cord neural progenitor	PDS kit: with modifications	EBSS	Mothe, 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
人	Human	神經(jīng)的，各種 / Neural, various	Papain: 12 u/ml； Trypsin: see reference； Collagenase/Dispase: see reference	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
人	Human, 9 month	神經(jīng)元 / Neurons	Collagenase Type 4: 1.33%； Papain: 0.07 u/ml； Neutral Protease: 1 mg/ml	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
人	Human, 26 week	神經(jīng)祖細(xì)胞 / Neural progenitor cells	Papain: 2.5 u/ml； Deoxyribonuclease I: 250 u/ml； Neutral Protease: 1 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
人	Human, adult	心室上皮 / Ventricular epithelial	Papain: 11.4 u/ml； Deoxyribonuclease I: 10 u/ml	DMEM/F12	Roy 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
人	Human, 5-65 years	視網(wǎng)膜色素上皮 (RPE) / 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	Human	背根神經(jīng)節(jié)神經(jīng)元 / Dorsal root ganglion neurons	Collagenase Type 1: 0.2%； Neutral Protease: 0.5%	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
昆蟲(chóng)	Drosophila, larval	神經(jīng)母細(xì)胞 / Neuroblasts	Collagenase Type 1: 0.1%； Papain: 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
昆蟲(chóng)	Drosophila	樹(shù)突 / Dendrites	Collagenase: 0.05%； Neutral Protease: 0.2%	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
昆蟲(chóng)	Gryllus Bimaculatus	巨大的中間神經(jīng)元 / Giant interneurons	Collagenase: 0.05%； Neutral Protease: 0.2%	Leibovitz's L15	Kloppenburg, 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