肝组织消化手册

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Worthington组织消化指南

引言

组织消化酶主要用于组织培养和细胞生物学研究的原代细胞的分离和收集。尽管这些消化酶已经普遍应用多年，但是对其组织消化和细胞收集的作用机制却没有全面的理解。由此导致的结果便是选择一种消化方法大多是随机选择或基于过去的经验，而不是在*理解消化原理的基础上选择的消化方案，以及如何根据具体实验目的修正消化方案以获得更好的实验结果。

细胞分离程序的目的是zui大程度的获得功能活性的单个细胞，而影响结果的原因有很多，包括（不限于）以下：

1、组织类型

2、物种来源

3、动物年龄

4、遗传改造情况（如knockouts）

5、所用的消化培养液

6、所用的酶

7、消化酶中的杂质

8、所用消化酶的工作浓度

9、温度

10、孵育时间

通常前四个原因没办法选择，而剩下的原因一般由经验决定以获取zui适实验结果。

通过查阅文献来选择*组织消化酶和zui适消化条件的研究人员通常被互相矛盾的实验数据搞得一头雾水。这些巨大差异来源于组织胞外基质的复杂而动态的特性，以及过去常用的粗制、未提纯的组织分离酶。另外，组织胞外基质由各种蛋白、糖蛋白、脂类和糖脂组成，这些成分的组成比例在不同物种、不同组织、甚至不同发育（年龄）之间存在明显的差异，这些因素都极大的影响了消化方案的制定。目前通常使用的粗制酶，如链霉蛋白酶、NF1:250、以及含有几种其他不同浓度蛋白酶的胶原酶、以及各种多糖酶、核酶和脂酶。

本指南总结了目前常用组织消化酶的zui适消化和细胞收集知识和方法，描述了标准的实验步骤，以便于制定一个*的细胞分离方案，同时列举了一些相关的参考文献。

下表列出多种组织的适宜消化酶（含参考文献，以组织类型和物种分类，详见链接）

脂肪组织	肾上腺组织	骨组织	脑组织
软骨组织	结肠组织	内皮组织	上皮组织
眼	心脏	肠	肾
肝	肺	淋巴结	乳腺
其它组织	其他组织	肌组织	神经组织
胰腺	腮腺	垂体	前列腺
生殖	动物鳞片	皮肤	脾脏
干细胞	胸腺	甲状腺/甲状旁腺	扁桃体
肿瘤

注：使用Worthington公司消化酶的文献不限于上表链接中所列，一般而言，Worthington公司的组织消化酶可用于大多组织类型的消化。

肝组织消化手册

物种	详细来源	细胞名称	消化用酶	培养基	参考文献
鸟类	鸟类, 各种各样的	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
鸟类	幼鸭, 7 天	Hepatocytes	Collagenase: 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
鸟类	幼鸭, 7 天	Hepatocytes	Hyaluronidase: 0.05%	DMEM/ F12
犬	狗, 13 岁大	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	(包括大鼠, 豚鼠, 兔, 人类)	Hepatocytes	Collagenase: 90 u/ml	CF EGTA perfusate
犬	狗, 成年的	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	鸡, 成年的	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	鸡	Hepatocytes	Collagenase Type 4: 6000 units	Medium A	Roseman, S., Weigel, P., Schnaar, R., Kuhlenschmidt, M., Schmell, E., Lee, R., and Lee, Y.: Adhesion of Hepatocytes to Immobilized Sugars. A Threshold Phenomenon, J Cell Biol 254, 10830, 1979
鸡	(包括 SD大鼠, 150-250 克)	Hepatocytes	Collagenase Type 4: 6000 units	Medium A
鸡	来亨鸡, 白色, 10-15 天	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	小鸡胚胎, 5 天	Heart	Trypsin: 3.0%	Tyrode's solution, CMF	Steinberg, M.: "ECM": Its Nature, Origin, And Function in Cell Aggregation, Exp Cell Res30, 257, 1963
鸡	小鸡胚胎, 5 天	Liver	Trypsin: 3.0%	Tyrode's solution, CMF
鸡	小鸡胚胎	Various tissues (heart, liver, skeletal, cardiac)	Trypsin: various grades	CMF Tyrode's solution	Rinaldini, L.: An Improved Method for the Isolation and Quantitative C*tion of Embryonic Cells, Exp Cell Res 16, 477, 1959
马	马, 4-13 年	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
鱼类	虹鳟鱼(虹鳟), 雄性 & 雌性,	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
鱼类	120-600 克	Hepatocytes	Collagenase Type 2: 0.045%	Kreb's-Ringer bicarbonate buffer, CF
鱼类	鲑鳟鱼, 雄性, 100 - 200 克	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
鱼类	虹鳟鱼, (Salmo 克airdneri), 雄性, 150-200 克	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
鱼类	鲑鳟鱼 100克- 5公斤	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
鱼类	鲑鳟鱼(虹鳟鱼), 雄性, 150-200 克	Hepatocytes	Collagenase: 100u/ml	HBSS	Klaunig, J.: Establishment of Fish Hepatocyte Cultures For Use in In VitroCarcinogenicity Studies, Natl Cancer Inst Monogr 65, 163, 1981
鱼类	鲶鱼(斑点叉尾鮰), 雄性, 100 - 150 克	Hepatocytes	Collagenase: 100u/ml	HBSS
豚鼠	Hartley豚鼠, , 雄性, 250-300 克	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	豚鼠, 年幼的	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
人类	人类, 胎儿	Liver Hematopoietic	Collagenase Type 4: 0.1%	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
			Hyaluronidase: 0.1%
			Deoxyribonuclease I: 2 u/ml
人类	人类	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
人类	人类	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
人类	人类	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
人类	人类	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
人类	人类	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
人类	人类, 成年人	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
人类	人类	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
人类	人类, 成年人	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
人类	人类	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
人类	(包括大鼠)	Hepatocytes	Collagenase Type 1: 0.05%	Leffert's buffer
人类	人类	Hepatocytes	Collagenase: 0.05%	Williams E	Donato M, Viitala P, Rodriguez-Antona C, Lindfors A, Casl 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
人类	人类	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
人类	人类	Hepatocytes	Collagenase: 0.05%	HEPES buffer	Gomez-Lechon, M., Lopez, P., Donato, T., Montoya, A., Larrauri, A., Gimenez, P., Trullenque, R., Fabra, R., and Casl, 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
人类	人类	Hepatocytes	Collagenase: 0.05%	(see reference)
人类	人类, 男性, 17-40 岁	Hepatocytes	Collagenase: 0.05%	HEPES buffer	Begue, J., Baffet, G., Campion, J., and Guillouzo, A.: Differential Response of Primary Cultures of Human and Rat Hepatocytes to Aflatoxin B1-Induced Cytotoxicity and Protection by the Hepatoprotective agent(+)-Cyanidanol-3, Biol Cell 63, 327, 1988
人类	人类, 4 男性肾捐献者, 年龄 17 到 31岁	Hepatocytes	Collagenase: 0.05% & 0.025%	HEPES	LeBot, M., Begue, J., Kernaleguen, D., Robert, J., Ratanasavanh, D., Airiau, J., Riche, C., and Guillouzo, A.: Different Cytotoxicity and Metabolism of Doxorubicin, Daunorubicin, Epirubicin, Esorubicin, and Idarubicin in Cultured Human and Rat Hepatocytes, Biochem Pharmacol 37 (20), 3877, 1988
人类	(包括成年大鼠, 雄性, SD, 180-200 克)	Hepatocytes	Collagenase: 0.05% & 0.025%	(see reference)
人类	人类, 3人肾移植, 年龄 20, 23, 25	Hepatocytes	Collagenase: 0.05%	HEPES	Gugen-Guillouzo, C., Campion, J., Brissot, P., Glaise, D., Launois, B., Bourel, M., and Guillouzo, A.: High Yield Preparation of Isolated Human Adult Hepatocytes by Enzymatic Perfusion of the Liver, Cell Biol Int Rep 6 (6), 625, 1982
人类	人类, 51-75 岁大	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
人类	(包括大鼠, 豚鼠, 兔,狗)	Hepatocytes	Collagenase: 90 u/ml	CF EGTA perfusate
人类	人类	Hepatocytes	Trypsin: 0.1%	HBSS, CMF	Kaighn, M.: Human Liver Cells,Tissue Culture Methods / Applications,(Kruse, P., Patterson, M. eds), , 54, 1973
猴	猴, 猕猴, 3-5.5 公斤	Hepatocytes	Collagenase Type 1: 129 u/ml	DMEM/F12	Weber, A., Groyer-Picard, M.and Dagher, I.: Hepatocyte Transplantation Techniques: Large Animal Models., Methods Mol Biol 481, 83, 2009
猴	猴 (猕猴), 成年的, 5-6 公斤	Hepatocytes	Trypsin: 160 u/ml	HEPES buffer	Ulrich, R., Aspar, D., Cramer, C., Kletzien, R., and Ginsberg, L.: Isolation and Culture of Hepatocytes from the Cynogmolgus Monkey (Macaca fascicularis), In Vitro Cell Dev Biol 26, 815, 1990
小鼠	小鼠	Hepatocytes	Collagenase Type 3: 100 u/ml	DMEM	Guenthner, C., Luitje, M., Pyle, L., Molyneux, P., Yu, J., Li, A., Leise, T. and Harrington, M.: Circadian Rhythms of Per2::Luc in Individual Primary Mouse Hepatocytes and Cultures., PLoS ONE 9, e87573, 2014
小鼠	小鼠	Hepatocytes	Collagenase Type 4: 100 u/ml	DMEM
小鼠	小鼠	hepatocytes	Collagenase Type 4: 100 u/ml	DMEM	Sin, Y., Ballantyne, L., Mukherjee, K., St Amand, T., Kyriakopoulou, L., Schulze, A. and Funk, C.: Inducible Arginase 1 Deficiency in Mice Leads to Hyperargininemia and Altered Amino Acid Metabolism., PLoS ONE 8, e80001, 2013
小鼠	小鼠	Hepatocytes & non-parenchymal liver cells	Collagenase: 0.03-0.05%	DMEM	Brundert, M., Heeren, J., Merkel, M., Carambia, A., Herkel, J., Groitl, P., Dobner, T., Ramakrishnan, R., Moore, K. and Rinninger, F.: Scavenger Receptor CD36 Mediates Uptake of High Density Lipoproteins in Mice and by Cultured Cells., J Lipid Res 52, 745, 2011
小鼠	小鼠	Hepatocytes	HIS kit: with modifications	DMEM	Kang, H., Okamoto, K., Kim, Y., Takeda, Y., Bortner, C., Dang, H., Wada, T., Xie, W., Yang, X., Liao, G. and Jetten, A.: Nuclear Orphan Receptor TAK1/TR4-Deficient Mice are Protected Against Obesity-Linked Inflammation, Hepatic Steatosis, and Insulin Resistance., Diabetes 60, 177, 2011
小鼠	小鼠	Hepatacytes	Collagenase Type 1: 0.05%	Williams E	Holl, D., Kuckenberg, P., Woynecki, T., Egert, A., Becker, A., Huss, S., Stabenow, D., Zimmer, A., Knolle, P., Tolba, R., Fischer, H. and Schorle, H.: Transgenic Overexpression of Tcfap2c/AP-2gamma Results in Liver Failure and Intestinal Dysplasia., PLoS ONE 6, e22034, 2011
小鼠	小鼠	CD133+ stem	Collagenase Type 4: 0.05%	DMEM/F12	Rountree, C., Ding, W., Dang, H., Vankirk, C. and Crooks, G.: Isolation of CD133+ Liver Stem Cells for Clonal Expansion., J Vis Exp 56, e3183, 2011
			Pronase: 0.05%
			Deoxyribonuclease I: 0.01%
小鼠	小鼠, 雄性, 12-14 周	Hepatocytes	Collagenase Type 1: 0.03%	Williams E	Chung, S., Timmins, J., Duong, M., Degirolamo, C., Rong, S., Sawyer, J., Singaraja, R., Hayden, M., Maeda, N., Rudel, L., Shelness, G. and Parks, J.: Targeted Deletion of Hepatocyte ABCA1 Leads to VLDL Triglyceride Over-Production and LDL Hypercatabolism, J Biol Chem 285, 12197, 2010
小鼠	小鼠	Liver non-parenchymal	Collagenase Type 3: 100 u/ml	HBSS	Bosschaerts, T., Guilliams, M., Stijlemans, B., Morias, Y., Engel, D., Tacke, F., Herin, M., De Baetselier, P. and Beschin, A.: Tip-DC Development During Parasitic Infection is Regulated by IL-10 and Requires CCL2/CCR2, IFN-gamma and MyD88 Signaling., PLoS Pathog 6, e1001045, 2010
小鼠	小鼠, 雄性, 成年的	Hepatocytes	Collagenase Type 4: 0.05%	HBSS	Mathijs, K., Kienhuis, A., Brauers, K. J, Jennen, D. , Lahoz, A., Kleinjans, J. and van Delft, J.: Assessing the Metabolic Competence of Sandwich-Cultured Mouse Primary Hepatocytes., Drug Metab Dispos 37, 1305, 2009
小鼠	小鼠, 雄性	Hepatocytes	Collagenase Type 1: 100 u/ml	Williams E	Oliva, J., Bardag-Gorce, F., French, B., Li, J., McPhaul, L., Amidi, F., Dedes, J., Habibi, A., Nguyen, S. and French, S.: Fat10 is an Epigenetic Marker for Liver Preneoplasia in a Drug-Primed Mouse Model of Tumorigenesis., Exp Mol Pathol 84, 102, 2008
小鼠	小鼠, 雄性	Hepatocytes	Elastase: 0.1 u/ml	Williams E
小鼠	小鼠, 7-10 周	Liver endothelial	Collagenase: 0.03%	DMEM	Follenzi, A., Benten, D., Novikoff, P., Faulkner, L., Raut, S. and Gupta, S.:Transplanted Endothelial Cells Repopulate the Liver Endothelium and Correct the Phenotype of Hemophilia A Mice., J Clin Invest 118, 935, 2008
小鼠	小鼠, 10-12 周	Liver sinusoidal endothelial cells	Collagenase Type 1: 0.05%	HEPES	Beldi, G., Wu, Y., Sun, X., Imai, M., Enjyoji, K., Csizmadia, E., Candinas, D., Erb, L. and Robson, S.: Regulated Catalysis of Extracellular Nucleotides by Vascular CD39/ENTPD1 is Required for Liver Regeneration., Gastroenterol 135, 1751, 2008
小鼠	小鼠, 10-12 周	Liver sinusoidal endothelial cells	Neutral Protease: 0.025%	HEPES
小鼠	小鼠	Liver epithelial progenitor cells	Collagenase Type 4: 0.1%	DMEM	Li Wen-Lin, Su Juan, Yao Yu-Cheng, Tao Xin-Rong, Yan Yong-Bi, Yu Hong-Yu, Wang Xin-Min, Li Jian-Xiu, Yang Yong-Ji, Lau Joseph T Y, Hu Yi-Ping: Isolation and characterization of bipotent liver progenitor cells from adult mouse, Stem Cells 24, 322-32, 2006
小鼠	小鼠	Liver epithelial progenitor cells	Deoxyribonuclease I: 0.05%	DMEM
小鼠	小鼠	Liver sinusodial endothelial	Collagenase: 0.03%	DMEM	Benten Daniel, Follenzi Antonia, Bhargava Kuldeep K, Kumaran Vinay, Palestro Christopher J, Gupta Sanjeev: Hepatic targeting of transplanted liver sinusoidal endothelial cells in intact mice, Hepatol 42, 140-8, 2005
小鼠	小鼠	Hepatocytes	Collagenase Type 1: 0.033%	Leffert's buffer	Jiang Guoqiang, Li Zhihua, Liu Franklin, Ellsworth Kenneth, Dallas-Yang Qing, Wu Margaret, Ronan John, Esau Christine, Murphy Cain, Szalkowski Deborah, Bergeron Raynald, Doebber Thomas, Zhang Bei: Prevention of obesity in mice by antisense oligonucleotide inhibitors of stearoyl-CoA desaturase-1, J Clin Invest 115, 1030-8, 2005
小鼠	小鼠, 8-10 周	Liver derived stem cells	Collagenase Type 1: 0.1%	HBSS	Kotton, D., Fabian, A. and Mulligan, R.: A Novel Stem-Cell Population in Adult Liver with Potent Hematopoietic-Reconstitution Activity., Blood 106, 1574, 2005
小鼠	小鼠, 8-10 周	Liver derived stem cells	Neutral Protease: 2.4 u/ml	HBSS
小鼠	小鼠	Hepatocytes	Collagenase Type 1: 0.053%	DMEM/ F-12	Sazani P., Gemignani F., Kang S., Maier M., Manoharan M., Persmark M., Bortner D., and Kole R.: Systemically Delivered Antisense Oligomers Up-regulate Gene Expression in Mouse Tissues, Nat Biotechnol 20, 1228, 2002
小鼠	小鼠, 雄性, 4-10 周	Hepatocytes	Collagenase Type 2: 100 u/ml	HBSS	Lingohr Melissa K, Bull Richard J, Kato-Weinstein Junko, Thrall Brian D:Dichloroacetate stimulates glycogen accumulation in primary hepatocytes through an insulin-independent mechanism, Toxicol Sci 68, 508-15, 2002
小鼠	小鼠, 雄性, 8 周	Hepatocytes	Collagenase Type 1: 0.04%	Waymouth's medium	Hatano E, Brenner DA: Akt protects mouse hepatocytes from TNF-alpha- and Fas-mediated apoptosis through NK-kappa B activation., Am J Physiol/Gastro 282, G1357, 2002
小鼠	小鼠	Hepatocytes	Collagenase: 0.05%	Williams E	Donato M, Viitala P, Rodriguez-Antona C, Lindfors A, Casl 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
小鼠	小鼠, 7 周	Kupffer cells	Collagenase Type 4: 0.05%	HBSS	Angele MK, Knoferl MW, Schwacha MG, Ayala A, Cioffi WG, Bland KI, and Chaudry IH.: Sex steroids regulate pro- and anti-inflammatory cytokine release by macrophages after trauma-hemorrhage, Am J Physiol 277, C35, 1999
小鼠	小鼠	Nonparenchymal liver	Collagenase Type 1: 0.05%	Hanks	Ling W, Lougheed M, Suzuki H, Buchan A, Kodama T, Steinbrecher UP: Oxidized or acetylated low density lipoproteins are rapidly cleared by the liver in mice with disruption of the scavenger receptor class A type I/II gene, J Clin Invest 100, 244-52, 1997
小鼠	小鼠, C3H, 6-8 周s	Epithelial	Collagenase: 0.10%	DMEM	Lillehaug, J., Mondal, S., and Heidelberger, C.: Establishment of Epithelial Cell Lines from Mouse Regenerating Liver, In Vitro 15, 910, 1979
小鼠	小鼠, 20-30 克	Parenchymal and non-parenchymal	Hyaluronidase: 0.1%	Hank's w/ Insulin, CMF	Crisp, D., and Pogson, C.: Glycolytic and Gluconeogenic Enzyme Activities in Parenchymal and Non-parenchymal Cells from Mouse Liver, Biochem J 126, 1009, 1972
猪	猪, 小型猪, 13 公斤	Hepatocytes	Collagenase Type 4: 0.05%	Williams E	Meng, F., Chen, Z., Han, M., Hu, X., He, X., Liu, Y., He, W., Huang, W., Guo, H. and Zhou, P.: Porcine Hepatocyte Isolation and Reversible Immortalization Mediated by Retroviral Transfer and Site-Specific Recombination., World J Gastroenterol 16, 1660, 2010
			Neutral Protease: 0.84%
			Deoxyribonuclease I: see reference
猪	约克郡猪, 雄性, 21天	Hepatocytes	Collagenase Type 1: 0.07%	Williams E	Terner, M., Gilmore, W.J., Lou, Y. and Squires, E.J.: The Role of CYP2A and CYP2E1 in the Metabolism of 3-Methylindole in Primary Cultured Porcine Hepatocytes., Drug Metab Dispos 34, 848, 2006
猪	猪, 中国小型猪, 6-10 公斤	Hepatocytes	Collagenase Type 4: 125 u/ml	Williams E	Li, J, Li, L., Chao, H., Yang, Q., Liu, X., Sheng, J., Yu, H. and Huang, J: Isolation and Short Term C*tion of Swine Hepatocytes for Bioartificial Liver Support System.,Hepatobiliary Pancreat Dis Int 4, 249, 2005
猪	猪, 1 周	Hepatocytes	Collagenase Type 4: 0.05%	RPMI 1640	Wang Y., Liu H., Guo H., Wen H., and Liu J.: Primary Hepatocyte Culture in Collagen Gel Mixture and Collagen Sandwich, World J Gastroenterol 10, 699, 2004
猪	猪, 雄性, 2-3 周	Hepatocytes	Collagenase: 0.05%	DMEM	Raman Priya, Donkin Shawn S, Spurlock Michael E: Regulation of hepatic glucose metabolism by leptin in pig and rat primary hepatocyte cultures, Am J Physiol Regul Integr Comp Physiol 286, R206-16, 2004
猪	猪, 成年的	Hepatocytes	Collagenase: 0.05%	MEM	Zhou X, Liu L, Kano J, Mukaiyama T, and Tokiwa T.: Isolation and C*tion of Porcine Hepatocytes for Extracorporeal Artificial Liver Support System, Chin Med J 114, 946, 2001
猪	猪, 成年的	Hepatocytes	Neutral Protease:	MEM
猪	猪, 雄性, 6-40 公斤	Hepatocytes	Collagenase: 0.8%	PBS	Gerlach, J., Brombacher, J., Smith, M., Neuhaus, P.: High Yield Hepatocyte Isolation from Pig Livers for Investigation of Hybrid Liver Support Systems: Influence of Collagenase Concentration and Body Weight, J Surg Res 62 (1), 85, 1996
兔	兔, 新西兰白	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
大鼠	SD大鼠, 500-700克	Hepatocytes and sinusoidal endothelial	Collagenase Type 4: 0.05%	HBSS	Perepelyuk, M., Terajima, M., Wang, A., Georges, P., Janmey, P., Yamauchi, M. and Wells, R.: Hepatic Slate Cells and Portal Fibroblasts are the Major Cellular Sources of Collagens and Lysyl Oxidases in Normal Liver and Early After Injury., Am J Physiol Gastrointest Liver Physiol 304, G605, 2013
大鼠	SD大鼠, 500-700克	Hepatocytes and sinusoidal endothelial	Deoxyribonuclease I: 0.003%	HBSS
大鼠	大鼠, 成年的	Hepatocytes	Collagenase Type 2: 1000 U	William's	Shen, L., Hillebrand, A., Wang, D. and Liu, M.: Isolation and Primary Culture of Rat Hepatic Cells., J Vis Exp 64, e3917, 2012
大鼠	大鼠, 成年的	Portal fibroblasts	Collagenase Type 2: 0.3%	DMEM/F-12	Wen, J., Olsen, A., Perepelyuk, M. and Wells, R.: Isolation of Rat Portal Fibroblasts by In Situ Liver Perfusion., J Vis Exp 64, e3669, 2012
大鼠	大鼠	Hepatocytes	Collagenase Type 2: 0.05%	DMEM	Budick-Harmelin, N., Anavi, S., Madar, Z. and Tirosh, O.: Fatty Acids-Stress Attenuates Gluconeogenesis Induction and Glucose Production in Primary Hepatocytes., Lipids Health Dis 11, 66, 2012
大鼠	SD大鼠, 240-320克	Hepatocytes	HIS kit: per instructions	Waymouth's MB	Pillai, V., and Mehvar, R.: Inhibition of NADPH-Cytochrome P450 Reductase by Tannic Acid in Rat Liver Microsomes and Primary Hepatocytes: Methodological Artifacts and Application to Ischemia-Reperfusion Injury, J Pharm Sci 100, 3495, 2011
大鼠	大鼠	Hepatocytes and SEC's	Collagenase Type 1: 0.1-0.2%	RPMI	Gopalakrishnan, S. and Harris, E.: In Vivo Liver Endocytosis Followed by Purification of Liver Cells by Liver Perfusion., J Vis Exp 57, e3138, 2011
大鼠	Wistar大鼠, 雌性	Hepatic slate cells	Collagenase Type 1: 0.025-0.1%	DMEM	Zvibel, I., Atias, D., Phillips, A., Halpern, Z. and Oren, R.: Thyroid Hormones Induce Activation of Rat Hepatic Slate Cells Through Increased Expression of p75 Neurotrophin Receptor and Direct Activation of Rho., Lab Invest 90, 674, 2010
大鼠	Wistar大鼠, 雌性	Hepatic slate cells	Pronase: 0.025-0.13%	DMEM
大鼠	SD大鼠	Hepatic slate cells	Pronase: 0.02%	Medium 199	Handy, J., Saxena, N., Fu, P., Lin, S., Mells, J., Gupta, N. and Anania, F.:Adiponectin Activation of AMPK Disrupts Leptin-Mediated Hepatic Fibrosis via Suppressors of Cytokine Signaling (SOCS-3)., J Cell Biochem 110, 1195, 2010
大鼠	SD大鼠	Hepatic slate cells	Collagenase: see reference	Medium 199
大鼠	SD大鼠, 250-300克	Hepatocytes	HIS kit: per instructions	Krebs-Henseleit	Parasrampuria, R. and Mehvar, R.: Dose-Dependent Inhibition of Transporter-Mediated Hepatic Uptake and Biliary Excretion of Methotrexate by Cyclosporine A in an Isolated Perfused Rat Liver Model., J Pharm Sci 99, 5060, 2010
大鼠	SD大鼠	Hepatocytes	Collagenase: 0.05%	HBSS	Chung, C., Shugrue, C., Nagar, A., Doll, J., Cornwell, M., Gattu, A., Kolodecik, T., Pandol, S. and Gorelick, F.: Ethanol Exposure Depletes Hepatic Pigment Epithelium-Derived Factor, a Novel Lipid Regulator., Gastroenterol 136, 331, 2009
大鼠	Lewis大鼠, 雄性	Hepatocytes	Collagenase Type 2: 0.1%	DPBS	Bettinger, C., Kulig, K., Vacanti, J., Langer, R. and Borenstein, J.: Nanofabricated Collagen-Inspired Synthetic Elastomers for Primary Rat Hepatocyte Culture., Tissue Eng Part A 15, 1321, 2009
大鼠	大鼠, 200-300 克	Hepatocytes	Collagenase Type 1: 0.067%	HBSS	Mula, N., Cubero, F., Codesal, J., de Andres, S., Escudero, C., Garcia-Barrutia, S., Millan, I., Arahuetes, R. and Maganto, P.: Survival of Allogeneic Hepatocytes Transplanted into the Thymus., Cells Tissues Organs 188, 270, 2008
大鼠	SD大鼠, 雄性, 230-250克	Hepatocytes	Collagenase Type 2: 0.1%	HEPES	Doleh, L. and Romani, A.: Biphasic Effect of Extra-Reticular Mg2+ on Hepatic G6P Transport and Hydrolysis., Arch Biochem Biophys 467, 283, 2007
大鼠	Lewis大鼠, 150-200克	Hepatocytes	Collagenase Type 2: 0.05%	Williams E	Smith, M. and Mooney, D.: Hypoxia Leads to Necrotic Hepatocyte Death., J Biomed Mater Res A 80, 520, 2007
大鼠	SD大鼠, 180-200克	Hepatocytes	Collagenase Type 1: 0.1%	see reference	Charbonneau, A., Unson, C. and Lavoie, J.: High-Fat Diet-Induced Hepatic Steatosis Reduces Glucagon Receptor Content in Rat Hepatocytes: Potential Interaction with Acute Exercise., J Physiol 579, 255, 2007
大鼠	Wistar大鼠, 270-330克	Hepatocytes	Collagenase Type 2: 0.033%	Williams E	Li, W., Ralphs, K., Slack, J. and Tosh, D.: Keratinocyte Serum-Free Medium Maintains Long-Term Liver Gene Expression and Function in Cultured Rat Hepatocytes by Preventing the Loss of Liver-Enriched Transcription Factors., Int J Biochem Cell Biol 39, 541-54, 2007
大鼠	SD大鼠, 500-750 克	Portal Fibroblasts	Collagenase: 0.03%	DMEM/F-12	Li, Z., Dranoff, J., Chan, E., Uemura, M., Sevigny, J.and Wells, R.: Transforming Growth Factor-beta and Substrate Stiffness Regulate Portal Fibroblast Activation in Culture., Hepatol 46, 1246-56, 2007
			Pronase: 0.033%
			Hyaluronidase: 0.036%
大鼠	大鼠,食鱼动物, E14	Hepatocytes	Collagenase Type 1: 0.22%	HBSS	Isabel Zvibel, Miri Bronstein, Einav Hubel, Ella Bar-Lev, Zamir Halpern, Ran Oren:Isolation, characterization and culture of Thy1-positive cells from fetal rat livers, World J Gastroenterol 12, 3841-7, 2006
大鼠	Lewis大鼠, 雌性, 150-200 克	Hepatocytes	Collagenase: see references	DMEM	Sosef MN, Baust JM, Sugimachi K, Fowler A, Tompkins RG, and Toner M.:Cryopreservation of isolated primary rat hepatocytes: enhanced survival and long-term hepatospecific function, Ann Surg 241, 125, 2005
大鼠	大鼠	Liver sinusodial endothelial	Collagenase: 0.03%	DMEM	Benten Daniel, Follenzi Antonia, Bhargava Kuldeep K, Kumaran Vinay, Palestro Christopher J, Gupta Sanjeev: Hepatic targeting of transplanted liver sinusoidal endothelial cells in intact mice, Hepatol 42, 140-8, 2005
大鼠	Wistar大鼠, 200-300 克	Hepatocytes	Collagenase Type 2: 0.05%	Williams E	Putz G, Schmider W, Nitschke R, Kurz G, Blum HE: Synthesis of phospholipid-conjugated bile salts and interaction of bile salt-coated liposomes with cultured hepatocytes, J Lipid Res 46, 2325-38, 2005
大鼠	大鼠, 成年的	Hepatocytes	Collagenase Type 1: 0.1%	Williams E	Jensen C., Jauho E., Santoni-Rugiu E., Holmskov U., Teisner B., Tygstrup N., and Bisgaard H.: Transit-amplifying Ductular (oval) Cells and Their Hepatocytic Progeny are Characterized by a Novel and Distinctive Expression of Delta-like Protein/preadipocyte Factor 1/fetal Antigen 1, Am J Pathol 164(4), 1347, 2004
			Pronase: 1%
			Deoxyribonuclease I: 0.007%
大鼠	SD大鼠, 雄性, 200-250 克	Hepatocytes	Collagenase: 0.05%	DMEM	Raman Priya, Donkin Shawn S, Spurlock Michael E: Regulation of hepatic glucose metabolism by leptin in pig and rat primary hepatocyte cultures, Am J Physiol Regul Integr Comp Physiol 286, R206-16, 2004
大鼠	Wistar大鼠	Hepatocytes	Collagenase Type 4: 0.05%	RPMI 1640	Wang Y., Liu H., Guo H., Wen H., and Liu J.: Primary Hepatocyte Culture in Collagen Gel Mixture and Collagen Sandwich, World J Gastroenterol 10, 699, 2004
大鼠	Wistar大鼠, 220-270克	Hepatocytes	Collagenase Type 1: 200 u/ml	DMEM	Annaert, P., Turncliff, R., Booth, C., Thakker, D. and Brouwer, K.: P-Glycoprotein-Mediated In Vitro Biliary Excretion in Sandwich-Cultured Rat Hepatocytes., Drug Metab Dispos 29, 1277-83, 2001
大鼠	SD大鼠, 雄性	Hepatocytes	Collagenase Type 2: 0.025%	Williams E	Burczynski, M., McMillian, M., Parker, J., Bryant, S., Leone, A., Grant, E., Thorne, J., Zhong, Z., Zivin, R. and Johnson, M.: Cytochrome P450 Induction in Rat Hepatocytes Assessed by Quantitative Real-Time Reverse-Transcription Polymerase Chain Reaction and the RNA Invasive Cleavage Assay., Drug Metab Dispos 29, 1243, 2001
大鼠	SD大鼠, 雄性, 230-250 克	Hepatocytes	Collagenase Type 1: 0.1%	HBSS	Kuddus, R., Nalesnik, M., Subbotin, V., Rao, A., and Gandhi, C.: Enhanced Sythesis and Reduced Metabolism of Endothelin-1 (ET-1) by Hepatocytes - An Important Mechanism of Increased Endogenous Levels of ET-1 in Liver Cirrhosis, J Hepatol 33, 725, 2000
大鼠	大鼠, 雄性, 3 周	Hepatocytes	Collagenase Type 4: 0.02%	Williams E	Low-Baselli A., Hufnagl K., Parzefall W., Schulte-Hermann R., and Grasl-Kraupp B.: Initiated Rat Hepatocytes in Primary Culture: A Novel Tool to Study Alterations in Growth Control During the First Stage of Carcinogenesis, Carcinogenesis 21, 79, 2000
大鼠	SD大鼠, 雄性, 450-500 克	Slate	Protease: 0.02%	HBSS	Gabriel, A., Kuddus, R., Rao, A., and Gandhi, C.: Down-Regulation of Endothelin Receptors by Transforming Growth Factor B1 in Hepatic Slate Cells, J Hepatol 30, 440, 1999
大鼠	Wistar大鼠, 雄性	Hepatocytes	Collagenase Type 2: 120 u/ml	HBSS	Wolz E, Liechti H, Notter B, Oesterhelt G, Kistler A: Characterization of metabolites of astaxanthin in primary cultures of rat hepatocytes, Drug Metab Dispos 27, 456-62, 1999
大鼠	Lewis大鼠, 成年的	Hepatocytes	Collagenase: 60 u/ml	HEPES	Matsuura T, Gad MZ, Harrison EH, and Ross AC: Lecithin:retinol acyltransferase and retinyl ester hydrolase activities are differentially regulated by retinoids and have distinct distributions between hepatocyte and nonparenchymal cell fractions of rat liver, J Nutr 127, 218, 1997
大鼠	大鼠	Hepatocytes	Collagenase: 0.04% - 0.06%	PBS	Alston-Smith, J and Pertoft, H: Isolation of Liver Cells: a System for Obtaining Pure Cells in Monolayer Cultures from a Single Rat Liver,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12B:14.1, 1995
大鼠	大鼠	Hepatocytes	Collagenase: 0.05%	DMEM	Matsumoto, K and Nakamura, T: Techniques for the Isolation and C*tion of Hepatocytes using Collagenase,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12B:16.1, 1995
大鼠	SD大鼠, 雄性, 200 克	Hepatocytes	Collagenase: 0.05%	Medium 199	Rana, B., Mischoulon, D., Xie, Y., Bucher, N., and Farmer, S.: Cell-Extracellular Matrix Interactions Can Regulate the Switch between Growth and Differentiation in Rat Hepatocytes: Reciprocal Expression of C/EBPa and Immediate-Early Growth Response Transcription Factors, Mol Cell Biol 14 (9), 5858, 1994
大鼠	Wistar大鼠, 成年的, 雄性, 100-125 克	Hepatocytes	Collagenase Type 2: 0.05%	Serum-free medium	Liu, J., McKim, J., Liu, Y., and Klaassen, C.: Effects of Butyrate Homologes on Metallothionein Induction in Rat Primary Hepatocyte Cultures, In Vitro Cell Dev Biol 28, 320, 1992
大鼠	SD大鼠, 雄性, 250-350 克	Hepatocytes	Collagenase Type 4: 200 u/ml	RPMI 1640	Dixit, V., Arthur, M., and Gitnick, G.: Repeated Transplantation of Microencapsulated Hepatocytes for Sustained Correction of Hyperbilirubinemia in Gunn Rats, Cell Transplant 1, 275, 1992
大鼠	大鼠,食鱼动物	Epithelial	Collagenase: 0.05%	Eagles's MEM	Blouin, M., and Marceau, N.: Primary Culture of Fetal Rat Liver Bipotential Progenitor Cells, J Tiss Cul Meth 13, 117, 1991
			Neutral Protease: 0.48%
			Hyaluronidase: 0.075%
大鼠	SD大鼠, 8 - 12 天	Parenchymal hepatocytes	Collagenase Type 4: 80 u/ml	HBSS	Davila, J., Reddy, C., Davis, P. and Acosta, D.: Toxicity Assessment of Papaverine Hydrochloride and Papaverine-Derived Metabolites in Primary Cultures of Rat Hepatocytes, In Vitro Cell Dev Biol 26, 515, 1990
大鼠	SD大鼠, 雄性, 250-350 克	Hepatocytes	Collagenase Type 2: 0.05%	HBSS, CMF	Liu, J., Kershaw, W., and Klaassen, C.: Rat Primary Hepatocyte Cultures are a Good Model for Examining Metallothionein-Induced Tolerance to Cadmium Toxicity, In Vitro Cell Dev Biol 26, 75, 1990
大鼠	Wistar大鼠, 雄性, 200 克	Parenchymal	Collagenase Type 1:	HEPES, modified	Kindberg, G., Gudmundsen, O., and Berg, T.: The Effect of Vandate on Receptor-mediated Endocytosis of Asialoorosomucoid in Rat Liver Parenchymal Cells, J Biol Chem 265, 8999, 1990
大鼠	SD大鼠, 雄性, 175-225 克	Hepatocytes	Collagenase:	Kreb's Ringer bicarbonate buffer	Reddy, S., Amick, G., Cooper, R., and Damun, Z.: Insulin Stimulates the Activity of a Protamine in Isolated Rat Hepatocytes, J Biol Chem 265, 7748, 1990
大鼠	SD大鼠, neonatal, 8 - 10 天	Hepatocytes	Collagenase Type 4: 0.05%	Hanks' BSS, CF	Davila, J., Lenherr, A., and Acosta, D.: Protective Effect of Flavonoids on Drug-Induced Hepatotoxicity In Vitro, Toxicology 57, 267, 1989
大鼠	大鼠,食鱼动物, 雄性, 125-250 克	Hepatocytes	Collagenase Type 1: 0.5%	RPMI 1640	Chan, K., Kost, D., and Michalopoulos, G.: Multiple Sequential Periods of DNA Synthesis and Quiescence in Primary Hepatocyte Cultures Maintained on the DMSO-EGF On/Off Protocol, J Cell Physiol 141, 584, 1989
大鼠	大鼠, 食鱼动物 344, 雄性, 14 个月	Hepatocytes	Collagenase: 0.05%	Ringer's biocarbonate buffer	Nagy, I., Ohno-Iwashita, Y., Ohta, M., Nagy, V., Kitani, K., Ando, S., and Imahori, K.: Effect of Perfringolysin O on the Lateral Diffusion Constant of Membrane Proteins of Hepatocytes as Revealed by Fluorescence Recovery After Photobleaching, Biochim Biophys Acta 939, 551, 1988
大鼠	SD大鼠, 雄性, 200-300 克	Hepatocytes	Collagenase: 0.05%	HEPES	Voss, A. and Sprecher, H.: Metabolism of 6,9,12-Octadecatetraenoic and 6,9,12,15-Octadecatetraenoic Acid, Biochim Biophys Acta 958, 153, 1988
大鼠	Wistar大鼠, 雄性,3 month, 200 克	Parenchymal	Collagenase Type 1: 0.05%	Krebs Henseleit	Kuiper, J., Zijlstra, F., Kamps, J, and vanBerkel, T.: Identification of Prostaglandin D2 as the Major Eiconsanoid from Liver Endothelial and Kupffer Cells, Biochim Biophys Acta 959, 143, 1988
		Endothelial
		Kupffer
大鼠	Wistar大鼠, 雄性, 200 - 300 克	Parenchymal	Collagenase Type 2: 0.05%	HBSS with CaCl₂	Cai, H., He, Z., and Ding, Y.: Effects of Monocyte Macrophages Stimulation on Hepatic Lipoprotein Receptors, Biochim Biophys Acta 958, 334, 1988
大鼠	Wistar大鼠, 雄性, 200 - 300 克	Kupffer	Collagenase Type 2: 0.05%	HBSS with CaCl₂
大鼠	Wistar大鼠, 雌性, 17 天	Hepatocytes	Collagenase: 0.05%	HBSS	Cotariu, D., Barr-Nea, L., Papo, N., and Zaidman, J.: Induction of gamma-Glutamyl Transferase by Dexamethasone in Cultured Rat Hepatocytes, Enzyme 40, 212, 1988
大鼠	大鼠, 镉应变, 白化变种, 雄性, 140 -180 克	Hepatocytes and Nonparenchymal	Pronase: 0.1%	HEPES buffer with calcium	Braun, L., Mead, J., Panzica, M., Mikumo, R., Bell, G., and Fausto, N.: Transforming Growth Factor B mRNA Increases During Liver Regeneration:A Possible Paracrine Mechanism Of Growth Regulation, Proc Natl Acad Sci U S A 85, 1539, 1988
大鼠	大鼠, 250-350 克	Hepatocytes	Collagenase Type 2: 166 u/ml	DMEM	Chang, T., and Chang, C.: Hepatic Uptake of Asialoglycoprotein is Different Among Mammalian Species Due to Different Receptor Distribution, Biochim Biophys Acta942, 57, 1988
大鼠	(包括豚鼠和兔)	Hepatocytes	Collagenase Type 2: 166 u/ml	DMEM
大鼠	SD大鼠, 雄性, 180-200 克	Hepatocytes	Collagenase: 0.05%	MEM	Gupta, C., Hattori, A., Betschart, J., Virji, M., and Shinozuka, H.: Modulation of Epidermal Growth Factor Receptors in Rat Hepatocytes by Two Liver-Promoting Regimens, a Choline-Deficient and a Phenobarbital Diet, Cancer Res 48, 1162, 1988
大鼠	大鼠,食鱼动物 344, 150-200 克	Bile ductular epithelial	Collagenase Type 1: 220 u/ml	L-15	Mathis, G., Walls, S., and Sirica, A.: Biochemical Characteristics of Hyperplastic Rat Bile Ductular Epithelial Cells Cultured "on Top" and "Inside" Different Extracellular Matrix Substitutes, Cancer Res 48, 6145, 1988
大鼠	Wistar大鼠, 雄性, 260-310 克	Hepatocytes	Collagenase Type 1: 0.05%	HEPES	McAbee DD and Weigel PH: ATP-dependent inactivation and reactivation of constitutively recycling galactosyl receptors in isolated rat hepatocytes,Biochemistry 27, 2061, 1988
大鼠	SD大鼠, 雄性, 350-400 克	Hepatocytes	Collagenase Type 2:	HBSS	Rodriguez de Turco, E., and Spitzer, J.: Kinetics of Diacylglycerol Accumulation in Response to Vasopressin Stimulation in Hepatocytes of Continuously Endotoxaemic Rats, Biochem J 253, 73, 1988
大鼠	SD大鼠, 雄性, 350 - 450 克	Lipocytes	Collagenase: 0.015%	DMEM/Ham's F-12	Friedman, S. and Roll, F.: Isolation and Culture of Hepatic Lipocytes,Kupffer Cells, and Sinusoidal Endothelial Cells by Density Gradient Centrifugation with Stractan, Anal Biochem 161, 207, 1987
		Kupffer
		Sinusoidal endothelial
大鼠	SD大鼠, 雄性	Hepatocytes	Collagenase: 100 - 200 礸/g body weight	Eagle's	Oka, J. and Weigel, P.: Monensin Inhibits Ligand Dissociation Only Transiently and Partially and distinguishes two galactosyl receptor pathways in isolated rat hepatocytes, J Cell Physiol 133, 243, 1987
				Eagle's w/HEPES
				HBSS
大鼠	大鼠, 食鱼动物, 雄性, 150-250 克	Hepatocytes	Collagenase Type 1: 125 - 250 u/ml	MEM	Francavilla, A., Ove, P., Polimeno, L., Sciascia, C., Coetzee, M., and Starzi, T.:Epidermal Growth Factor and Proliferation in Rat Hepatocytes in Primary Culture Isolation at Different Times after Partial Hepatectomy, Cancer Res 46, 1318, 1986
大鼠	大鼠, 食鱼动物, 雄性, 150-250 克	Hepatocytes	Collagenase Type 1: 125 - 250 u/ml	(see reference)
大鼠	SD大鼠, 雄性, 200-250 克	Hepatocytes	Collagenase: 0.05%	HBSS	Kreamer, B., Staecker, J., Sawada, N., Sattler, G., Hsia, M., and Pitot, H.: Use of Low-Speed, Iso-Density Percoll Centrifugation Method to Increase the Viability of Isolated Rat Hepatocyte Preparations, In Vitro Cell Dev Biol 22 (4), 201, 1986
大鼠	SD大鼠, 雌性, 100-180 克	Hepatocytes	Collagenase: 0.5%	Krebs Ringer bicarbonate buffer	Schwarz, K., Lanier, S., Carter, E., Homcy, C., and Graham, R.: Rapid Reciprocal Changes in Adrenergic Receptors in Intact Isolated Hepatocytes During Primary Cell Culture, Mol Pharmacol 27, 200, 1985
大鼠	大鼠, Lewis, 雌性, 200-250 克	Hepatocytes, Kupffer, endothelial	Collagenase: 0.05%	Gey's BSS	Holstege A, Leser HG, Pausch J, Gerok W: Uridine catabolism in Kupffer cells, endothelial cells, and hepatocytes, Eur J Biochem 149, 169-73, 1985
大鼠	SD大鼠, 雄性, 270 - 320克	Hepatocytes	Collagenase: 0.04%	Bicarbonate buffer with calcium added	Brass, E., Garrity, M., and Robertson, R.: Inhibition of Glucagon-Stimulated Hepatic Glycogenolysis by E-Series Prostaglandins, F.E.B.S. Lett. 169, 293, 1984
大鼠	Wistar大鼠, 成年的, 雄性, 150-200 克	Hepatocytes	Trypsin: 0.005%	Williams E	Okumura, T. and Saito, K.: Degradation of Prostaglandin E2 in a Primary Culture of 成年的 Rat Hepatocytes, J. Biochem. 96, 429, 1984
大鼠	大鼠, 食鱼动物, Lewis and SD, 雄性 10-18 months	Epithelial	Trypsin: 0.05%	HBSS CMF	Herring, A., Raychaudhuri, R., Kelley, S., and Iybe, P.: Repeated Establishment of Diploid Epithelial Cell Cultures from Normal and Partially Hepatectomized Rats, In Vitro 19, 576, 1983
大鼠	Wistar大鼠, 雄性, 3 month old	Endothelial	Collagenase: 0.05%	HBSS	Nagelkenke, J., Barto, K., and Berkel, T.: In Vivo and in Vitro Uptake and Degradation of Acetylated Low Density Lipoprotein by Rat Liver Endothelial, Kupffer, and Parenchymal Cells, J Biol Chem 258 (20), 12221, 1983
		Kupffer
		Parenchymal
大鼠	SD大鼠, 雄性/雌性, 250-300 克	Hepatocytes	Collagenase: 100 u/ml	Krebs Henseleit bicarbonate buffer	Studer, R. and Borle, A.: Differences between Male and Female Rats in the Regulation of Hepatic Glycogenolysis. The Relative Role of Calcium and cAMP in Phosphorylase Activation by Catecholamines, J Biol Chem 257, 7987, 1982
大鼠	Wistar大鼠, 12 天	Epithelial	Trypsin: 0.05%	HBSS, CMF	Malan-Shibley, L., and Iype, P.: Influence of Cultures on Cell Morphology/Tyrosine Aminotransferase Levels, Exp Cell Res 131, 363, 1981
大鼠	Zucker大鼠, 过度肥胖	Hepatocytes	Collagenase Type 2: 0.30%	Dulbecco-Vogt arginine free Eagle's	Goldstein, A., Palmer, J., and Johnson, P.: Primary Cultures of Fetal Hepatocytes from the Genetically Obese Zucker Rat: Protein Synthesis, In Vitro 17, 651, 1981
大鼠	SD大鼠	Hepatocytes	Collagenase: 90 u/ml	CF EGTA perfusate	Reese, J. and Byard, J.: Isolation And Culture of 成年的 Hepatocytes from Liver Biopsies, In Vitro 17, 935, 1981
大鼠	Wistar大鼠, 雌性, fetus	Hepatocytes	Collagenase: 0.025%	HEPES buffer	Gugen-Guillouzo, C., Tichonicky, L., Szajnert, M., and Kruh, J.: Changes in Some Chromatin and Cytoplastic Enzymes of Perinatal Rat Hepatocytes, In Vitro 16, 1, 1980
大鼠	SD大鼠, 雄性, 250-300 克	Parenchymal	Collagenase Type 2: 0.05%	Kreb's Henseleit bicarbonate buffer	Yamada, S., Otto, P., Kennedy, D., and Whayne, T.: The Effects of Dexamethasone on Metabolic Activity of Hepatocytes in Primary Monolayer Culture, In Vitro 16, 559, 1980
大鼠	Wistar大鼠, 雄性, 180-210 克	Hepatocytes	Hyaluronidase: 0.02%	Kreb's buffer	De Gerlache, J., Lans, M., Taper, H., and Roberfroid, M.: Separate Isolation of Cells from Nodules and Surrounding Parenchyma of the Same Precancerous Rat Liver: Biochemical and Cytochemical Characterization, Toxicology 18, 225, 1980
大鼠	SD大鼠, 7-10 天	Liver	Collagenase: 0.05%	HBSS modified (see reference)	Acosta, D., Anuforo, D., and Smith, R: J Tiss Cul Meth 6, 35, 1980
大鼠	Wistar大鼠, 雄性, 200-250 克	Hepatocytes	Hyaluronidase: 460 u/ml	Saline	Poli, G., Gravela, E., Albano, E., and Dianzani, M.: Studies on Fatty Liver with Isolated Hepatocytes: II., The Action of Carbon Tetrachloride on Lipid Peroxidation, Protein and Triglyceride Synthesis and Secretion, Exp Mol Pathol 30, 116, 1979
大鼠	SD大鼠, 雄性, 100-150 克	Hepatocytes	Collagenase Type 1: 0.065%	DMEM	Davis, R., Engelhorn, S., Pangburn, S., Weinstein, D., and Steinberg, D.: Very Low Density Lipoprotein Synthesis and Secretion by Cultured Rat Hepatocytes, J Biol Chem254 (6), 2010, 1979
大鼠	SD大鼠, 雄性, 100-150 克	Hepatocytes	Collagenase Type 1: 0.065%	(see reference)
大鼠	大鼠	Hepatocytes	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
大鼠	(also chicken)	Hepatocytes	Collagenase Type 3 & 4:	HEPES
大鼠	SD大鼠, 雄性, 200 克	Hepatocytes	Collagenase Type 1: 100 u/ml	Buffers 1 & 2	Rubin, K., Kjellen, L., and Oslashbrink, B.: Intercellular Adhesion between Juvenile Liver Cells.. A Method to Measure the Formation of Stable Lateral Contacts Between Cells Attached to a Collagen Gel, Exp Cell Res 109, 413, 1977
大鼠	SD大鼠, 雄性, 200 克	Hepatocytes	Collagenase Type 1: 100 u/ml	(see reference)
大鼠	大鼠 (WAG/RIJ), 雌性, 24, 30, 和 37 个月	Parenchymal	Collagenase Type 1: 0.05% - 0.06%	HEPES buffer	VanBezodijen, C., Grell, T., and Knook, D.: Effect of Age on Protein Synthesis by Isolated Liver Parenchymal Cells, Mech Ageing Dev 6, 293, 1977
大鼠	大鼠,食鱼动物, 成年的, 雄性	Hepatocytes	Collagenase Type 1: 100 u/ml	HBSS	Williams, G., Bermudez, E., and Scaramuzzino, D.: Rat Hepatocytes Primary Cell Cultures III. Improved Dissociation and Attachment Techniques and the Enchancement of Survival by Culture Medium, In Vitro 13 (12), 809, 1977
大鼠	大鼠,食鱼动物, 成年的, 雄性	Hepatocytes	Collagenase Type 1: 100 u/ml	(see reference)
大鼠	Wistar大鼠 , 雄性, 250-300 克	Hepatocytes	Collagenase: 0.05%	Hank's solution, CF	Gravela, E., Poli, G., Albano, E., and Dianzani, M.: Studies of Fatty Liver with Isolated Hepatocytes, Exp Mol Pathol 27, 339, 1977
大鼠	大鼠, 食鱼动物, 雄性, 成年的, 170-265 克	Hepatocytes	Collagenase: 0.05%-0.10%	Williams E	Laishes, B., and Williams, G.: Conditions Affecting Primary Cell Cultures of Functional 成年的 Rat Hepatocytes. I.. The Effect of Insulin, In Vitro 12, 521, 1976
大鼠	SD大鼠, 雄性, 200-250 克	Hepatocytes	Collagenase Type 1: 0.05%	MEM	Witters, L., Alberico, L., and Acruch, J.: Insulin Regulation of Glycogen Synthase in the Isolated Rat Hepatocyte, Biochem Biophys Res Commun 69 (4), 997, 1976
大鼠	大鼠, 食鱼动物, 成年的, 200-250 克	Epithelial	Hyaluronidase: 0.0075%	KCl-NaCl HEPES Buffer	Williams, G., and Gunn, J.: Long-Term Culture of 成年的 Rat Liver Epithelial Cells, Exp Cell Res 89, 139, 1974
大鼠	大鼠, 初生, 3 天	Hepatocytes	Trypsin: 0.25%	PBS, CMF	Bausher, J., and Schaeffer, W.: A Diploid Rat Liver Cell Culture. 1. Characterization and Sensitivity to Aflatoxin B1, In Vitro 9, 286, 1974
大鼠	SD大鼠, 白化变种, 雄性	Parenchymal	Hyaluronidase: 0.1%	Hank's solution, CMF	Bonney, R., Becker, J., Walker, P., and Potter, V.: Primary Monolayer Cultures of 成年的 Rat Liver Parenchymal Cells Suitable for Study of the Regulation of Enzyme Synthesis, In Vitro 9, 399, 1974
大鼠	大鼠, 胚胎, 1-3 天s	Hepatocytes	Hyaluronidase: 0.10%	HBSS	Gerschenson, L., Berliner, J., and Davidson, M.: The Isolation and Culture of Liver Cells, Vol. 32,, , 733, 1974
大鼠	SD大鼠, 成年的, 雄性, 200-250 克	Parenchymal	Hyaluronidase: 0.10%	HBSS, CF	Howard, R., Lee, J., and Pesch, L.: The fine structure, potassium content, and respiratory activity of isolated rat liver parenchymal cells prepared by improved enzymatic techniques, J Cell Biol 57, 642, 1973
大鼠	SD大鼠, 成年的, 雄性, 180-300 克	Parenchymal	Collagenase Type 1: 0.05%	Hank's solution, CF	Bissell, D., Hammaker, L., and Meyer, U.: Parenchymal Cells from 成年的 Rat Liver in Nonproliferating Monolayer Culture. I. Functional Studies, J Cell Biol 59, 722, 1973
大鼠	Wistar大鼠, 200-250 克	Hepatocytes	Hyaluronidase: 0.10%	HBSS, CF	Berg, T., Boman, D., and Seglen, P.O.: Induction of Tryptophan Oxygenase in Primary Rat Liver Cell Suspensions by Glucocorticoid Hormone, Exp Cell Res 72, 571, 1972
大鼠	SD大鼠, 雌性, 130-160 克	Hepatocytes	Hyaluronidase: 0.08%	HBSS, CF	Johnson, M., Das, N., Butcher, F., and Fain, J.: The Regulation of Gluconeogenesis in Isolated Rat Liver Cells by Glucagon, Insulin, Dibutyryl Cyclic Adenosine Monophosphate, and Fatty Acids, J Biol Chem 247, 3229, 1972
大鼠	大鼠,食鱼动物 344, 怀孕的 (19-21 天妊娠期),	Hepatocytes	Collagenase: 0.3%	Modified Eagle's w/ Serum	Leffert, H., and Paul, D.: Studies on Primary Cultures of Differentiated Fetal Liver Cells, J Cell Biol 52, 559, 1972
大鼠	200-250 克	Hepatocytes	Collagenase: 0.3%	Modified Eagle's w/ Serum
大鼠	Wistar大鼠, 雄性, 260-310 克	Hepatocytes	Collagenase: 0.01 - 0.08%	HEPES	Seglen, P.: Preparation of Rat Liver Cells, Exp Cell Res 74, 450, 1972
大鼠	大鼠,食鱼动物, 10 天	Epithelial-like	Trypsin: 0.25%	PBS	Williams, G., Weisburger, E., and Weisburger, J.: Isolation and Long-Term Cell Culture of Epithelial-Like Cells from Rat Liver, Exp Cell Res 69, 106, 1971
大鼠	Wistar大鼠, 雄性, 6-8 周, 80-160 克	Hepatocytes	Hyaluronidase: 1.0%	HBSS, CMF	Iype, P.: Cultures from 成年的 Rat Liver Cells. 1.Establishment of Monolayer Cell-Cultures from Normal Lver, J Cell Physiol 78, 281, 1971
大鼠	大鼠, 100-200 克	Hepatocytes	Hyaluronidase: 0.10%	HBSS, CF	Haung, Y., and Ebner, K.: Induction of Tyrosine Aminotransferase in Isolated Liver Cells, Biochim Biophys Acta 191, 161, 1969
大鼠	SD大鼠, 成年的, 200-300 克	Parenchymal	Hyaluronidase: 0.10%	HBSS, CF	Berry, M., and Friend, D.: High Yield Preparation of Isolated Rat Liver Parenchymal Cells, J Cell Biol 43, 506, 1969
大鼠	SD大鼠, 成年的, 200-300 克	Parenchymal	Collagenase Type 1: 0.05%	HBSS, CF

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