Worthington 肝脏组织分离指南

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，针对肝脏细胞组织涉及到肝细胞，肝边缘群，肝细胞和血窦内皮，肝脏非实质，各种组织（心脏，肝脏，骨骼，心脏）等等的解离通常会使用酶，如何选择合适的酶来完成对应肝脏细胞组织的解离，以及原代细胞采集的常规操作， 艾美捷针对肝脏细胞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：肝脏组织分离指南

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。

肝脏
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	Avian, various	肝细胞 / Hepatocytes	Collagenase Type 1: 0.1%	see reference	Else, P., Brand, M., Turner, N. and Hulbert, A: Respiration Rate of Hepatocytes Varies With Body Mass in Birds, J Exp Biol 207, 2305, 2004
禽类	Ducklings, 7 day	肝细胞 / Hepatocytes	Collagenase: 0.05%； Hyaluronidase: 0.05%	DMEM/ F12	Lee J., Culvenor J., Angus P., Smallwood R., Nicoll A., and Locarnini S.: Duck Hepatitis B Virus Replication in Primary Bile Duct Epithelial Cells, J Virol 75(16), 7651, 2001
犬/狗	Dog, 13 years old (also rat, guinea-pig, rabbit, human)	肝细胞 / Hepatocytes	Collagenase: 90 u/ml	CF EGTA perfusate	Reese, J. and Byard, J.: Isolation And Culture of Adult Hepatocytes from Liver Biopsies, In Vitro 17, 935, 1981
犬/狗	Dog, adult	肝细胞 / Hepatocytes	Trypsin: 0.1%	HBSS, CMF	Vickrey, H., Ramon, J., and McCann, D.: Continuous Culture of Normal Adult Mammalian Hepatocytes Exhibiting Parenchymal Functions, In Vitro 15, 120, 1979
鸡	Chicken, adult	肝细胞 / Hepatocytes	Collagenase: 0.02%	HEPES, CF	Fraslin, J., Touquette, L., Douaire, M., Menezo, Y., Guillemot, J., and Mallard, J.: Isolation and Long Term Maintenance of Differentiated Adult Chicken Hepatocytes in Primary Culture, In Vitro Cell Dev Biol 28, 615, 1992
鸡	Chicken (also rat, SD, 150-250 g)	肝细胞 / Hepatocytes	Collagenase Type 4: 6000 units； Collagenase Type 3 & 4:	HEPES	Weigel, P., Schnaar, R., Kuhlenschmidt, M., Schmell, E., Lee, R., Lee, Y., and Roseman, S.: Adhesion of Hepatocytes to Immobilized Sugars, J Biol Chem 254 (21), 10830, 1979
鸡	Chicken, Leghorn, white, 10-15 day	肝细胞 / Hepatocytes	Deoxyribonuclease I: 0.00125%	PBS	Tarlow, D., Watkins, P., Reed, R., Miller, R., Zwergel, E., and Lane, M.: Lipogenesis and the Synthesis and Secretion of Very Low Density Lipoprotein by Avian Liver Cells in Nonproliferating Monolayer Culture, J Cell Biol 73, 332, 1974
鸡	Chick embryos, 5 day	心，肝 / Heart，Liver	Trypsin: 3.0%	Tyrode's solution, CMF	Steinberg, M.: "ECM": Its Nature, Origin, And Function in Cell Aggregation, Exp Cell Res 30, 257, 1963
鸡	Chick embryonic	各种组织（心脏，肝脏，骨骼，心脏） / Various tissues (heart, liver, skeletal, cardiac)	Trypsin: various grades	CMF Tyrode's solution	Rinaldini, L.: An Improved Method for the Isolation and Quantitative Cultivation of Embryonic Cells, Exp Cell Res 16, 477, 1959
马	Equine, 4-13 yr	肝细胞 / Hepatocytes	Collagenase Type 4: 0.1%	HBSS	Bakala A., Karlik W., and Wiechetek M.: Preparation of Equine Isolated Hepatocytes, Toxicol In Vitro 17(5-6), 615, 2003
鱼	Rainbow trout (Oncorhynchus mykiss), male & female, 120-600 g	肝细胞 / Hepatocytes	Collagenase Type 2: 0.045%	Kreb's-Ringer bicarbonate buffer, CF	Blair, J.B., Miller, M.R., Pack, D., Barnes, R., Teh, S.J. and Hinton, D.E.: Isolated Trout Liver Cells:Establishing Short-Term Primary Cultures Exhibiting Cell-to-Cell Interactions, In Vitro Cell Dev Biol 26, 237, 1990
鱼	Trout, male, 100 - 200 g	肝细胞 / Hepatocytes	Collagenase Type 2: 0.045%	HBSS with 0.05M HEPES	Lipsky, M., Sheridan, T., Bennett, R., and May, E.: Comparison of Trout Hepatocyte Culture on Different Substrates, In Vitro Cell Dev Biol 22, 360, 1986
鱼	Rainbow trout, (Salmo gairdneri), male, 150-200 g	肝细胞 / Hepatocytes	Collagenase: 100 u/ml	HBSS/CMF	Klaunig, J., Ruch, R., and Goldblah, P.: Trout Hepatocyte Culture: Isolation and Primary Culture, In Vitro Cell Dev Biol 21, 221, 1985
鱼	Trout 100g - 5kg	肝细胞 / Hepatocytes	Hyaluronidase: 0.08%	See reference	Bailey, G., Taylor, M., and Selivonchick, D.: Aflatoxin B1 Metabolism and DNA Binding in Isolated Hepatocytes From Rainbow Trout Salmo gairdner, Carcinogenesis 3, 511, 1982
鱼	Trout (Salmo gairdneri), male, 150-200 g；Catfish (Ictalurus punctatus), male, 100 - 150 g	肝细胞 / Hepatocytes	Collagenase: 100u/ml	HBSS	Klaunig, J.: Establishment of Fish Hepatocyte Cultures For Use in In Vitro Carcinogenicity Studies, Natl Cancer Inst Monogr 65, 163, 1981
豚鼠	Guinea-pig, Hartley, male, 250-300 g	肝细胞 / Hepatocytes	Collagenase Type 2:	Kreb's Ringer bicarbonate buffer	Arinze, I. and Kawai, Y.: Adrenergic Regulation of Glycogenolysis in Isolated Guinea-Pig Hepatocytes: Evidence that B2-Receptors Mediate Catecholamine Stimulation of Glycogenolysis, Arch Biochem Biophys 225, 196, 1983
豚鼠	Guinea-pig, young	肝细胞 / Hepatocytes	Collagenase: 90 u/ml	CF EGTA perfusate	Reese, J. and Byard, J.: Isolation And Culture of Adult Hepatocytes from Liver Biopsies, In Vitro 17, 935, 1981
人	Human	肝细胞 / Hepatocytes	Collagenase Type 1: 0.05%	HBSS	Reuschel, E, Jilg, W., Seelbach-Goebel, B. and Demi, L.: Comparative Purification and Characterization of Hepatitis B Virus-like Particles Produced by Recombinant Vaccinia Viruses in Human Hepatoma Cells and Human Primary Hepatocytes, PLoS ONE , , 2019
人	Human	肝细胞 / Hepatocytes	Collagenase Type 4: 0.06%	DMEM	Werner, M., Driftmann, S., Kleinehr, K., Kaiser, G., Mathe, Z., Treckmann, J., Paul, A., Skibbe, K., Timm, J., Canbay, A., Gerken, G., Schlaak, J. and Broering, R.: All-In-One: Advanced Preparation of Human Parenchymal and Non-Parenchymal Liver Cells., PLoS ONE 10, e0138655, 2015
人	Human	肝细胞和非实质细胞 / Hepatocytes and non-parenchymal cells	Collagenase Type 1: 0.1%	HBSS	Pfeiffer, E., Kegel, V., Zeilinger, K., Hengstler, J., Nussler, A., Seehofer, D. and Damm, G.: Featured Article: Isolation, Characterization, and Cultivation of Human Hepatocytes and Non-Parenchymal Liver Cells., Exp Biol Med (Maywood) 240, 645-56, 2015
人	Human	肝细胞 / Hepatocytes	Collagenase: 0.1-0.15%	HEPES	Lee, S., Schelcher, C., Demmel, M., Hauner, M. and Thasler, W.: Isolation of Human Hepatocytes by a Two-Step Collagenase Perfusion Procedure., J Vis Exp , , 2013
人	Human, fetal	肝脏造血 / Liver hematopoietic	Collagenase Type 4: 0.1%； Hyaluronidase: 0.1%； Deoxyribonuclease I: 2 u/ml	RPMI	Vatakis, D., Bristol, G., Kim, S., Levin, B., Liu, W., Radu, C., Kitchen, S. and Zack, J.: Using the BLT Humanized Mouse as a Stem Cell Based Gene Therapy Tumor Model., J Vis Exp 70, e4181, 2012
人	Human	肝细胞 / Hepatocytes	Collagenase Type 1: 0.05%	Williams E	Torok, E., Lutgehetmann, M., Bierwolf, J., Melbeck, S., Dullmann, J., Nashan, B., Ma, P. and Pollok, J.: Primary Human Hepatocytes on Biodegradable Poly(l-lactic acid) Matrices: A Promising Model for Improving Transplantation Efficiency with Tissue Engineering., Liver Transpl 17, 104, 2011
人	Human	肝干细胞和成肝细胞 / Hepatic stem cells and heptoblasts	Collagenase Type 4: 0.014-0.06%	various	Wauthier, E., Schmelzer, E., Turner, W., Zhang, L., LeCluyse, E., Ruiz, J., Turner, R., Furth, M., Kubota, H., Lozoya, O., Barbier, C., McClelland, R., Yao, H., Moss, N., Bruce, A., Ludlow, J. and Reid, L.: Hepatic Stem Cells and Hepatoblasts: Identification, Isolation, and Ex Vivo Maintenance., Methods Cell Biol 86, 137, 2008
人	Human	肝细胞 / Hepatocytes	Collagenase: 0.05%	HEPES	Pichard L, Raulet E, Fabre G, Ferrini JB, Ourlin JC, and Maurel P: Human hepatocyte culture, Methods Mol Biol 320, 283, 2006
人	Human	肝细胞 / Hepatocytes	Collagenase: 0.05%	EBSS	Hughes, R., Mitry, R., Dhawan, A., Lehec, S., Girlanda, R., Rela, M., Heaton, N., and Muiesan, P.: Isolation of Hepatocytes from Livers from Non-Heart-Beating Donors for Cell Transplantation, Liver Transpl 12, 713, 2006
人	Human	肝边缘群 / Hepatic side population	Collagenase: 0.02-0.05%	HBSS	Hussain, S., Strom, S., Kirby, M., Burns, S., Langemeijer, S., Ueda, T., Hsieh, M. and Tisdale, J.: Side Population Cells Derived from Adult Human Liver Generate Hepatocyte-Like Cells In Vitro., Dig Dis Sci 50, 1755, 2005
人	Human, adult	肝细胞 / Hepatocytes	Collagenase: 0.6%	RPMI 1640	Cho JJ, Joseph B, Sappal BS, Giri RK, Wang R, Ludlow JW, Furth ME, Susick R, and Gupta S.: Analysis of the functional integrity of cryopreserved human liver cells including xenografting in immunodeficient mice to address suitability for clinical applications, Liver Int 24, 361, 2004
人	Human	肝细胞 / Hepatocytes	Collagenase Type 4: 0.05%	Williams E	Duanmu Z., Locke D., Smigelski J., Wu W., Dahn M., Falany C., Kocarek T., and Runge-Morris M.: Effects of Dexamethasone on Aryl (SULT1A1)- and hydroxysteroid (SULT2A1)-Sulfotransferase Gene Expression in Primary Cultured Human Hepatocytes, Drug Metab Dispos 30(9), 997, 2002
人	Human, fetal	上皮祖细胞 / Epithelial progenitor	Collagenase: 0.03%	DMEM	Malhi, H., Irani, A., Gagandeep, S. and Gupta, S.: Isolation of Human Progenitor Liver Epithelial Cells with Extensive Replication Capacity and Differentiation into Mature Hepatocytes., J Cell Sci 115, 2679, 2002
人	Human (also rat)	肝细胞 / Hepatocytes	Collagenase Type 1: 0.05%	Leffert's buffer	Dandri, M., Burda, M., Torok, E., Pollok, J., Iwanska, A., Sommer, G., et al.: Repopulation of Mouse Liver with Human Hepatocytes and In Vivo Infection with Hepatitis B Virus, Hepatol 33, 981, 2001
人	Human	肝细胞 / Hepatocytes	Collagenase: 0.05%	Williams E	Donato M, Viitala P, Rodriguez-Antona C, Lindfors A, Castell J, Raunio H, Gomez-Lechon M, Pelkonen O: CYP2A5/CYP2A6 expression in mouse and human hepatocytes treated with various in vivo inducers, Drug Metab Dispos 28, 1321-6, 2000
人	Human	肝细胞 / Hepatocytes	Collagenase Type 1: 0.025%	Williams E	Parzefall, W., Erber, E., Sedivy, R., and Schulte-Hermann, R.: Testing for Induction of DNA Synthesis in Human Hepatocyte Primary Cultures by Rat Liver Tumor Promoters, Cancer Res 51, 1143, 1991
人	Human	肝细胞 / Hepatocytes	Collagenase: 0.05%	HEPES buffer (see reference)	Gomez-Lechon, M., Lopez, P., Donato, T., Montoya, A., Larrauri, A., Gimenez, P., Trullenque, R., Fabra, R., and Castell, J.: Culture of Human Hepatocytes From Small Surgical Liver Biopsies: Biochemical Characterization And Comparison With in vivo, In Vitro Cell Dev Biol 26, 67, 1990