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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 / HepatocytesCollagenase Type 1: 0.1%see referenceElse, 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 / HepatocytesCollagenase: 0.05%;
Hyaluronidase: 0.05%
DMEM/ F12Lee 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 / HepatocytesCollagenase: 90 u/mlCF EGTA perfusateReese, 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 / HepatocytesTrypsin: 0.1%HBSS, CMFVickrey, H., Ramon, J., and McCann, D.: Continuous Culture of Normal Adult Mammalian Hepatocytes Exhibiting Parenchymal Functions, In Vitro 15, 120, 1979
Chicken, adult肝细胞 / HepatocytesCollagenase: 0.02%HEPES, CFFraslin, 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
Chicken (also rat, SD, 150-250 g)肝细胞 / HepatocytesCollagenase Type 4: 6000 units;
Collagenase Type 3 & 4:
HEPESWeigel, 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
Chicken, Leghorn, white, 10-15 day肝细胞 / HepatocytesDeoxyribonuclease I: 0.00125%PBSTarlow, 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
Chick embryos, 5 day心,肝 / Heart,LiverTrypsin: 3.0%Tyrode's solution, CMFSteinberg, M.: "ECM": Its Nature, Origin, And Function in Cell Aggregation, Exp Cell Res 30, 257, 1963
Chick embryonic各种组织(心脏,肝脏,骨骼,心脏) / Various tissues (heart, liver, skeletal, cardiac)Trypsin: various gradesCMF Tyrode's solutionRinaldini, L.: An Improved Method for the Isolation and Quantitative Cultivation of Embryonic Cells, Exp Cell Res 16, 477, 1959
Equine, 4-13 yr肝细胞 / HepatocytesCollagenase Type 4: 0.1%HBSSBakala A., Karlik W., and Wiechetek M.: Preparation of Equine Isolated Hepatocytes, Toxicol In Vitro 17(5-6), 615, 2003
Rainbow trout (Oncorhynchus mykiss), male & female, 120-600 g肝细胞 / HepatocytesCollagenase Type 2: 0.045%Kreb's-Ringer bicarbonate buffer, CFBlair, 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
Trout, male, 100 - 200 g肝细胞 / HepatocytesCollagenase Type 2: 0.045%HBSS with 0.05M HEPESLipsky, M., Sheridan, T., Bennett, R., and May, E.: Comparison of Trout Hepatocyte Culture on Different Substrates, In Vitro Cell Dev Biol 22, 360, 1986
Rainbow trout, (Salmo gairdneri), male, 150-200 g肝细胞 / HepatocytesCollagenase: 100 u/mlHBSS/CMFKlaunig, J., Ruch, R., and Goldblah, P.: Trout Hepatocyte Culture: Isolation and Primary Culture, In Vitro Cell Dev Biol 21, 221, 1985
Trout 100g - 5kg肝细胞 / HepatocytesHyaluronidase: 0.08%See referenceBailey, G., Taylor, M., and Selivonchick, D.: Aflatoxin B1 Metabolism and DNA Binding in Isolated Hepatocytes From Rainbow Trout Salmo gairdner, Carcinogenesis 3, 511, 1982
Trout (Salmo gairdneri), male, 150-200 g;Catfish (Ictalurus punctatus), male, 100 - 150 g肝细胞 / HepatocytesCollagenase: 100u/mlHBSSKlaunig, 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 / HepatocytesCollagenase Type 2:Kreb's Ringer bicarbonate bufferArinze, 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 / HepatocytesCollagenase: 90 u/mlCF EGTA perfusateReese, J. and Byard, J.: Isolation And Culture of Adult Hepatocytes from Liver Biopsies, In Vitro 17, 935, 1981
Human肝细胞 / HepatocytesCollagenase Type 1: 0.05%HBSSReuschel, 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
Human肝细胞 / HepatocytesCollagenase Type 4: 0.06%DMEMWerner, 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
Human肝细胞和非实质细胞 / Hepatocytes and non-parenchymal cellsCollagenase Type 1: 0.1%HBSSPfeiffer, 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
Human肝细胞 / HepatocytesCollagenase: 0.1-0.15%HEPESLee, 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
Human, fetal肝脏造血 / Liver hematopoieticCollagenase Type 4: 0.1%;
Hyaluronidase: 0.1%;
Deoxyribonuclease I: 2 u/ml
RPMIVatakis, 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
Human肝细胞 / HepatocytesCollagenase Type 1: 0.05%Williams ETorok, 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
Human肝干细胞和成肝细胞 / Hepatic stem cells and heptoblastsCollagenase Type 4: 0.014-0.06%variousWauthier, 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
Human肝细胞 / HepatocytesCollagenase: 0.05%HEPESPichard L, Raulet E, Fabre G, Ferrini JB, Ourlin JC, and Maurel P: Human hepatocyte culture, Methods Mol Biol 320, 283, 2006
Human肝细胞 / HepatocytesCollagenase: 0.05%EBSSHughes, 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
Human肝边缘群 / Hepatic side populationCollagenase: 0.02-0.05%HBSSHussain, 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
Human, adult肝细胞 / HepatocytesCollagenase: 0.6%RPMI 1640Cho 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
Human肝细胞 / HepatocytesCollagenase Type 4: 0.05%Williams EDuanmu 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
Human, fetal上皮祖细胞 / Epithelial progenitorCollagenase: 0.03%DMEMMalhi, 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
Human (also rat)肝细胞 / HepatocytesCollagenase Type 1: 0.05%Leffert's bufferDandri, 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
Human肝细胞 / HepatocytesCollagenase: 0.05%Williams EDonato 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
Human肝细胞 / HepatocytesCollagenase Type 1: 0.025%Williams EParzefall, 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
Human肝细胞 / HepatocytesCollagenase: 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


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