Worthington 肝臟組織分離指南

發(fā)布者：艾美捷科技發(fā)布時間：2022-06-07

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原代細胞分離中，針對肝臟細胞組織涉及到肝細胞，肝邊緣群，肝細胞和血竇內(nèi)皮，肝臟非實質(zhì)，各種組織（心臟，肝臟，骨骼，心臟）等等的解離通常會使用酶，如何選擇合適的酶來完成對應(yīng)肝臟細胞組織的解離，以及原代細胞采集的常規(guī)操作， 艾美捷針對肝臟細胞相關(guān)的組織分類解離分種屬、不同的細胞、酶組合推薦，推薦濃度、培養(yǎng)基以及參考文獻，詳細匯總指南：肝臟組織分離指南

本技術(shù)指南描述了標準實驗室程序；為建立細胞分離方案提供了一種合乎邏輯的實驗方法；并列出了許多組織特異性參考資料，以幫助開發(fā)有效的方法。更多其他組織的解離指南，點擊查看：各組織分離指南。

肝臟
種屬	種屬詳情	細胞名稱	酶組合推薦，含推薦濃度	培養(yǎng)基	參考文獻
禽類	Avian, various	肝細胞 / 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
禽類	Ducklings, 7 day	肝細胞 / 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)	肝細胞 / 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	肝細胞 / 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
雞	Chicken, adult	肝細胞 / 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
雞	Chicken (also rat, SD, 150-250 g)	肝細胞 / 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
雞	Chicken, Leghorn, white, 10-15 day	肝細胞 / 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
雞	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
雞	Chick embryonic	各種組織（心臟，肝臟，骨骼，心臟） / 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
馬	Equine, 4-13 yr	肝細胞 / 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
魚	Rainbow trout (Oncorhynchus mykiss), male & female, 120-600 g	肝細胞 / 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
魚	Trout, male, 100 - 200 g	肝細胞 / 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
魚	Rainbow trout, (Salmo gairdneri), male, 150-200 g	肝細胞 / 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
魚	Trout 100g - 5kg	肝細胞 / 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
魚	Trout (Salmo gairdneri), male, 150-200 g；Catfish (Ictalurus punctatus), male, 100 - 150 g	肝細胞 / 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	肝細胞 / 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	肝細胞 / 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	肝細胞 / 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	肝細胞 / 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	肝細胞和非實質(zhì)細胞 / 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	肝細胞 / 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	肝臟造血 / 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	肝細胞 / 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	肝干細胞和成肝細胞 / 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	肝細胞 / 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	肝細胞 / 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	肝邊緣群 / 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	肝細胞 / 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	肝細胞 / 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	上皮祖細胞 / 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)	肝細胞 / 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	肝細胞 / 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	肝細胞 / 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	肝細胞 / 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