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肝組織消化手冊(cè)

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Worthington組織消化指南

引言

組織消化酶主要用于組織培養(yǎng)和細(xì)胞生物學(xué)研究的原代細(xì)胞的分離和收集。盡管這些消化酶已經(jīng)普遍應(yīng)用多年,但是對(duì)其組織消化和細(xì)胞收集的作用機(jī)制卻沒有全面的理解。  由此導(dǎo)致的結(jié)果便是選擇一種消化方法大多是隨機(jī)選擇或基于過去的經(jīng)驗(yàn),而不是在*理解消化原理的基礎(chǔ)上選擇的消化方案,以及如何根據(jù)具體實(shí)驗(yàn)?zāi)康男拚桨敢垣@得更好的實(shí)驗(yàn)結(jié)果。

細(xì)胞分離程序的目的是zui大程度的獲得功能活性的單個(gè)細(xì)胞,而影響結(jié)果的原因有很多,包括(不限于)以下:

1、組織類型

2、物種來源

3、動(dòng)物年齡

4、遺傳改造情況(如knockouts

5、所用的消化培養(yǎng)液

6、所用的酶

7、消化酶中的雜質(zhì)

8、所用消化酶的工作濃度

9、溫度

10、孵育時(shí)間

通常前四個(gè)原因沒辦法選擇,而剩下的原因一般由經(jīng)驗(yàn)決定以獲取zui適實(shí)驗(yàn)結(jié)果。

通過查閱文獻(xiàn)來選擇*組織消化酶和zui適消化條件的研究人員通常被互相矛盾的實(shí)驗(yàn)數(shù)據(jù)搞得一頭霧水。這些巨大差異來源于組織胞外基質(zhì)的復(fù)雜而動(dòng)態(tài)的特性,以及過去常用的粗制、未提純的組織分離酶。另外,組織胞外基質(zhì)由各種蛋白、糖蛋白、脂類和糖脂組成,這些成分的組成比例在不同物種、不同組織、甚至不同發(fā)育(年齡)之間存在明顯的差異,這些因素都極大的影響了消化方案的制定。目前通常使用的粗制酶,如鏈霉蛋白酶、NF1:250、以及含有幾種其他不同濃度蛋白酶的膠原酶、以及各種多糖酶、核酶和脂酶。

本指南總結(jié)了目前常用組織消化酶的zui適消化和細(xì)胞收集知識(shí)和方法,描述了標(biāo)準(zhǔn)的實(shí)驗(yàn)步驟, 便于制定一個(gè)*的細(xì)胞分離方案,同時(shí)列舉了一些相關(guān)的參考文獻(xiàn)。

下表列出多種組織的適宜消化酶(含參考文獻(xiàn),以組織類型和物種分類,詳見鏈接)

脂肪組織

腎上腺組織

骨組織

腦組織

軟骨組織

結(jié)腸組織

內(nèi)皮組織

上皮組織

心臟

淋巴結(jié)

乳腺

其它組織

其他組織

肌組織

神經(jīng)組織

胰腺

腮腺

垂體

前列腺

生殖

動(dòng)物鱗片

皮膚

脾臟

干細(xì)胞

胸腺

甲狀腺/甲狀旁腺

扁桃體

腫瘤

 

 

 

注:使用Worthington公司消化酶的文獻(xiàn)不限于上表鏈接中所列,一般而言,Worthington公司的組織消化酶可用于大多組織類型的消化。

 

肝組織消化手冊(cè)

 

物種詳細(xì)來源細(xì)胞名稱消化用酶培養(yǎng)基參考文獻(xiàn)
鳥類鳥類, 各種各樣的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
鳥類幼鴨, 7 天HepatocytesCollagenase: 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
Hyaluronidase: 0.05%
狗, 13 歲大HepatocytesCollagenase: 90 u/mlCF EGTA perfusateReese, J. and Byard, J.: Isolation And Culture of Adult Hepatocytes from Liver Biopsies, In Vitro 17, 935, 1981
(包括大鼠, 豚鼠, 兔, 人類)
狗, 成年的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
雞, 成年的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
雞 HepatocytesCollagenase Type 4: 6000 unitsMedium ARoseman, 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
(包括 SD大鼠, 150-250 克)
來亨雞, 白色, 10-15 天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
小雞 胚胎, 5 天HeartTrypsin: 3.0%Tyrode's solution, CMFSteinberg, M.: "ECM": Its Nature, Origin, And Function in Cell Aggregation, Exp Cell Res30, 257, 1963
Liver
小雞 胚胎Various tissues (heart, liver, skeletal, cardiac)Trypsin: various gradesCMF Tyrode's solutionRinaldini, L.: An Improved Method for the Isolation and Quantitative C*tion of Embryonic Cells, Exp Cell Res 16, 477, 1959
馬, 4-13 年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
魚類虹鱒魚(虹鱒), 雄性 & 雌性, 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
120-600 克
魚類鮭鱒魚, 雄性, 100 - 200 克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
魚類虹鱒魚, (Salmo 克airdneri), 雄性, 150-200 克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
魚類鮭鱒魚 100克- 5公斤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
魚類鮭鱒魚(虹鱒魚), 雄性, 150-200 克HepatocytesCollagenase: 100u/mlHBSSKlaunig, J.: Establishment of Fish Hepatocyte Cultures For Use in In VitroCarcinogenicity Studies, Natl Cancer Inst Monogr 65, 163, 1981
鯰魚(斑點(diǎn)叉尾鮰), 雄性, 100 - 150 克
豚鼠Hartley豚鼠, , 雄性, 250-300 克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
豚鼠豚鼠, 年幼的HepatocytesCollagenase: 90 u/mlCF EGTA perfusateReese, J. and Byard, J.: Isolation And Culture of Adult Hepatocytes from Liver Biopsies, In Vitro 17, 935, 1981
人類人類, 胎兒Liver HematopoieticCollagenase Type 4: 0.1%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
Hyaluronidase: 0.1%
Deoxyribonuclease I: 2 u/ml
人類人類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
人類人類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
人類人類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
人類人類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
人類人類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
人類人類, 成年人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
人類人類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
人類人類, 成年人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
人類人類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
(包括大鼠)
人類人類HepatocytesCollagenase: 0.05%Williams EDonato 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
人類人類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
人類人類HepatocytesCollagenase: 0.05%HEPES bufferGomez-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
(see reference)
人類人類, 男性, 17-40 歲HepatocytesCollagenase: 0.05%HEPES bufferBegue, 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
人類人類, 4 男性腎捐獻(xiàn)者, 年齡 17 到 31歲HepatocytesCollagenase: 0.05% & 0.025%HEPESLeBot, 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
(包括 成年大鼠, 雄性, SD, 180-200 克)(see reference)
人類人類, 3人腎移植, 年齡 20, 23, 25HepatocytesCollagenase: 0.05%HEPESGugen-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
人類人類, 51-75 歲大HepatocytesCollagenase: 90 u/mlCF EGTA perfusateReese, J. and Byard, J.: Isolation And Culture of Adult Hepatocytes from Liver Biopsies, In Vitro 17, 935, 1981
(包括大鼠, 豚鼠, 兔,狗)
人類人類HepatocytesTrypsin: 0.1%HBSS, CMFKaighn, M.: Human Liver Cells,Tissue Culture Methods / Applications,(Kruse, P., Patterson, M. eds), , 54, 1973
猴, 獼猴, 3-5.5 公斤HepatocytesCollagenase Type 1: 129 u/mlDMEM/F12Weber, A., Groyer-Picard, M.and Dagher, I.: Hepatocyte Transplantation Techniques: Large Animal Models., Methods Mol Biol 481, 83, 2009
猴 (獼猴), 成年的, 5-6 公斤HepatocytesTrypsin: 160 u/mlHEPES bufferUlrich, 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HepatocytesCollagenase Type 3: 100 u/mlDMEMGuenthner, 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
Collagenase Type 4: 100 u/ml
小鼠小鼠hepatocytesCollagenase Type 4: 100 u/mlDMEMSin, 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 cellsCollagenase: 0.03-0.05%DMEMBrundert, 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HepatocytesHIS kit: with modificationsDMEMKang, 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HepatacytesCollagenase Type 1: 0.05%Williams EHoll, 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+ stemCollagenase Type 4: 0.05%DMEM/F12Rountree, 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HepatocytesCollagenase Type 1: 0.03%Williams EChung, 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-parenchymalCollagenase Type 3: 100 u/mlHBSSBosschaerts, 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HepatocytesCollagenase Type 4: 0.05%HBSSMathijs, 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HepatocytesCollagenase Type 1: 100 u/mlWilliams EOliva, 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
Elastase: 0.1 u/ml
小鼠小鼠, 7-10 周Liver endothelialCollagenase: 0.03%DMEMFollenzi, 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 cellsCollagenase Type 1: 0.05%HEPESBeldi, 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
Neutral Protease: 0.025%
小鼠小鼠Liver epithelial progenitor cellsCollagenase Type 4: 0.1%DMEMLi 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
Deoxyribonuclease I: 0.05%
小鼠小鼠Liver sinusodial endothelialCollagenase: 0.03%DMEMBenten 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HepatocytesCollagenase Type 1: 0.033%Leffert's bufferJiang 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 cellsCollagenase Type 1: 0.1%HBSSKotton, D., Fabian, A. and Mulligan, R.: A Novel Stem-Cell Population in Adult Liver with Potent Hematopoietic-Reconstitution Activity., Blood 106, 1574, 2005
Neutral Protease: 2.4 u/ml
小鼠小鼠HepatocytesCollagenase Type 1: 0.053%DMEM/ F-12Sazani 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HepatocytesCollagenase Type 2: 100 u/mlHBSSLingohr 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HepatocytesCollagenase Type 1: 0.04%Waymouth's mediumHatano 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HepatocytesCollagenase: 0.05%Williams EDonato 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 cellsCollagenase Type 4: 0.05%HBSSAngele 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 liverCollagenase Type 1: 0.05%HanksLing 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sEpithelialCollagenase: 0.10%DMEMLillehaug, 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-parenchymalHyaluronidase: 0.1%Hank's w/ Insulin, CMFCrisp, D., and Pogson, C.: Glycolytic and Gluconeogenic Enzyme Activities in Parenchymal and Non-parenchymal Cells from Mouse Liver, Biochem J 126, 1009, 1972
豬, 小型豬, 13 公斤HepatocytesCollagenase Type 4: 0.05%Williams EMeng, 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
約克郡豬, 雄性, 21天HepatocytesCollagenase Type 1: 0.07%Williams ETerner, 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
豬, 中國小型豬, 6-10 公斤HepatocytesCollagenase Type 4: 125 u/mlWilliams ELi, 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
豬, 1 周HepatocytesCollagenase Type 4: 0.05%RPMI 1640Wang 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
豬, 雄性, 2-3 周HepatocytesCollagenase: 0.05%DMEMRaman 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
豬, 成年的HepatocytesCollagenase: 0.05%MEMZhou 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
Neutral Protease: 
豬, 雄性, 6-40 公斤HepatocytesCollagenase: 0.8%PBSGerlach, 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, 新西蘭白HepatocytesCollagenase: 90 u/mlCF EGTA perfusateReese, 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 endothelialCollagenase Type 4: 0.05%HBSSPerepelyuk, 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
Deoxyribonuclease I: 0.003%
大鼠大鼠, 成年的HepatocytesCollagenase Type 2: 1000 UWilliam'sShen, 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 fibroblastsCollagenase Type 2: 0.3%DMEM/F-12Wen, 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HepatocytesCollagenase Type 2: 0.05%DMEMBudick-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HepatocytesHIS kit: per instructionsWaymouth's MBPillai, 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'sCollagenase Type 1: 0.1-0.2%RPMIGopalakrishnan, 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 cellsCollagenase Type 1: 0.025-0.1%DMEMZvibel, 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
Pronase: 0.025-0.13%
大鼠 SD大鼠Hepatic slate cellsPronase: 0.02%Medium 199Handy, 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
Collagenase: see reference
大鼠 SD大鼠, 250-300克HepatocytesHIS kit: per instructionsKrebs-HenseleitParasrampuria, 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HepatocytesCollagenase: 0.05%HBSSChung, 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HepatocytesCollagenase Type 2: 0.1%DPBSBettinger, 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HepatocytesCollagenase Type 1: 0.067%HBSSMula, 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HepatocytesCollagenase Type 2: 0.1%HEPESDoleh, 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HepatocytesCollagenase Type 2: 0.05%Williams ESmith, 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HepatocytesCollagenase Type 1: 0.1%see referenceCharbonneau, 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HepatocytesCollagenase Type 2: 0.033%Williams ELi, 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 FibroblastsCollagenase: 0.03%DMEM/F-12Li, 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,食魚動(dòng)物, E14HepatocytesCollagenase Type 1: 0.22%HBSSIsabel 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HepatocytesCollagenase: see referencesDMEMSosef 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 endothelialCollagenase: 0.03%DMEMBenten 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HepatocytesCollagenase Type 2: 0.05%Williams EPutz 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HepatocytesCollagenase Type 1: 0.1%Williams EJensen 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HepatocytesCollagenase: 0.05%DMEMRaman 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HepatocytesCollagenase Type 4: 0.05%RPMI 1640Wang 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HepatocytesCollagenase Type 1: 200 u/mlDMEMAnnaert, 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HepatocytesCollagenase Type 2: 0.025%Williams EBurczynski, 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HepatocytesCollagenase Type 1: 0.1%HBSSKuddus, 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HepatocytesCollagenase Type 4: 0.02%Williams ELow-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SlateProtease: 0.02%HBSSGabriel, 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HepatocytesCollagenase Type 2: 120 u/mlHBSSWolz 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HepatocytesCollagenase: 60 u/mlHEPESMatsuura 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HepatocytesCollagenase: 0.04% - 0.06%PBSAlston-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HepatocytesCollagenase: 0.05%DMEMMatsumoto, 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HepatocytesCollagenase: 0.05%Medium 199Rana, 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HepatocytesCollagenase Type 2: 0.05%Serum-free mediumLiu, 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HepatocytesCollagenase Type 4: 200 u/mlRPMI 1640Dixit, 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,食魚動(dòng)物EpithelialCollagenase: 0.05%Eagles's MEMBlouin, 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 hepatocytesCollagenase Type 4: 80 u/mlHBSSDavila, 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HepatocytesCollagenase Type 2: 0.05%HBSS, CMFLiu, 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ParenchymalCollagenase Type 1: HEPES, modifiedKindberg, 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HepatocytesCollagenase: Kreb's Ringer bicarbonate bufferReddy, 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HepatocytesCollagenase Type 4: 0.05%Hanks' BSS, CFDavila, J., Lenherr, A., and Acosta, D.: Protective Effect of Flavonoids on Drug-Induced Hepatotoxicity In Vitro, Toxicology 57, 267, 1989
大鼠大鼠,食魚動(dòng)物, 雄性, 125-250 克HepatocytesCollagenase Type 1: 0.5%RPMI 1640Chan, 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, 食魚動(dòng)物 344, 雄性, 14 個(gè)月HepatocytesCollagenase: 0.05%Ringer's biocarbonate bufferNagy, 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HepatocytesCollagenase: 0.05%HEPESVoss, 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ParenchymalCollagenase Type 1: 0.05%Krebs HenseleitKuiper, 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ParenchymalCollagenase Type 2: 0.05%HBSS with CaCl2Cai, H., He, Z., and Ding, Y.: Effects of Monocyte Macrophages Stimulation on Hepatic Lipoprotein Receptors, Biochim Biophys Acta 958, 334, 1988
Kupffer
大鼠Wistar大鼠, 雌性, 17 天HepatocytesCollagenase: 0.05%HBSSCotariu, 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, 鎘應(yīng)變, 白化變種, 雄性, 140 -180 克Hepatocytes and NonparenchymalPronase: 0.1%HEPES buffer with calciumBraun, 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HepatocytesCollagenase Type 2: 166 u/mlDMEMChang, T., and Chang, C.: Hepatic Uptake of Asialoglycoprotein is Different Among Mammalian Species Due to Different Receptor Distribution, Biochim Biophys Acta942, 57, 1988
(包括豚鼠和兔)
大鼠SD大鼠, 雄性, 180-200 克HepatocytesCollagenase: 0.05%MEMGupta, 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,食魚動(dòng)物 344, 150-200 克Bile ductular epithelialCollagenase Type 1: 220 u/mlL-15Mathis, 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HepatocytesCollagenase Type 1: 0.05%HEPESMcAbee 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HepatocytesCollagenase Type 2: HBSSRodriguez 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LipocytesCollagenase: 0.015%DMEM/Ham's F-12Friedman, 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HepatocytesCollagenase: 100 - 200 礸/g body weightEagle'sOka, 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, 食魚動(dòng)物, 雄性, 150-250 克HepatocytesCollagenase Type 1: 125 - 250 u/mlMEMFrancavilla, 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
(see reference)
大鼠SD大鼠, 雄性, 200-250 克HepatocytesCollagenase: 0.05%HBSSKreamer, 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HepatocytesCollagenase: 0.5%Krebs Ringer bicarbonate bufferSchwarz, 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, endothelialCollagenase: 0.05%Gey's BSSHolstege 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HepatocytesCollagenase: 0.04%Bicarbonate buffer with calcium addedBrass, 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HepatocytesTrypsin: 0.005%Williams EOkumura, 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, 食魚動(dòng)物, Lewis and SD, 雄性 10-18 monthsEpithelialTrypsin: 0.05%HBSS CMFHerring, 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 oldEndothelialCollagenase: 0.05%HBSSNagelkenke, 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HepatocytesCollagenase: 100 u/mlKrebs Henseleit bicarbonate bufferStuder, 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EpithelialTrypsin: 0.05%HBSS, CMFMalan-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, 過度肥胖HepatocytesCollagenase Type 2: 0.30%Dulbecco-Vogt arginine free Eagle'sGoldstein, 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HepatocytesCollagenase: 90 u/mlCF EGTA perfusateReese, 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, fetusHepatocytesCollagenase: 0.025%HEPES bufferGugen-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ParenchymalCollagenase Type 2: 0.05%Kreb's Henseleit bicarbonate bufferYamada, 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HepatocytesHyaluronidase: 0.02%Kreb's bufferDe 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LiverCollagenase: 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HepatocytesHyaluronidase: 460 u/mlSalinePoli, 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HepatocytesCollagenase Type 1: 0.065%DMEMDavis, 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
(see reference)
大鼠大鼠HepatocytesCollagenase 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
(also chicken)
大鼠SD大鼠, 雄性, 200 克HepatocytesCollagenase Type 1: 100 u/mlBuffers 1 & 2Rubin, 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
(see reference)
大鼠大鼠 (WAG/RIJ), 雌性, 24, 30, 和 37 個(gè)月ParenchymalCollagenase Type 1: 0.05% - 0.06%HEPES bufferVanBezodijen, 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,食魚動(dòng)物, 成年的, 雄性HepatocytesCollagenase Type 1: 100 u/mlHBSSWilliams, 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
(see reference)
大鼠Wistar大鼠 , 雄性, 250-300 克HepatocytesCollagenase: 0.05%Hank's solution, CFGravela, 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, 食魚動(dòng)物, 雄性, 成年的, 170-265 克HepatocytesCollagenase: 0.05%-0.10%Williams ELaishes, 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HepatocytesCollagenase Type 1: 0.05%MEMWitters, 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, 食魚動(dòng)物, 成年的, 200-250 克EpithelialHyaluronidase: 0.0075%KCl-NaCl HEPES BufferWilliams, 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HepatocytesTrypsin: 0.25%PBS, CMFBausher, 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, 白化變種, 雄性ParenchymalHyaluronidase: 0.1%Hank's solution, CMFBonney, 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sHepatocytesHyaluronidase: 0.10%HBSSGerschenson, 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ParenchymalHyaluronidase: 0.10%HBSS, CFHoward, 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ParenchymalCollagenase Type 1: 0.05%Hank's solution, CFBissell, 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HepatocytesHyaluronidase: 0.10%HBSS, CFBerg, 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HepatocytesHyaluronidase: 0.08%HBSS, CFJohnson, 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,食魚動(dòng)物 344, 懷孕的 (19-21 天妊娠期), HepatocytesCollagenase: 0.3%Modified Eagle's w/ SerumLeffert, H., and Paul, D.: Studies on Primary Cultures of Differentiated Fetal Liver Cells, J Cell Biol 52, 559, 1972
200-250 克
大鼠Wistar大鼠, 雄性, 260-310 克HepatocytesCollagenase: 0.01 - 0.08%HEPESSeglen, P.: Preparation of Rat Liver Cells, Exp Cell Res 74, 450, 1972
大鼠大鼠,食魚動(dòng)物, 10 天Epithelial-likeTrypsin: 0.25%PBSWilliams, 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HepatocytesHyaluronidase: 1.0%HBSS, CMFIype, 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HepatocytesHyaluronidase: 0.10%HBSS, CFHaung, 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ParenchymalHyaluronidase: 0.10%HBSS, CFBerry, M., and Friend, D.: High Yield Preparation of Isolated Rat Liver Parenchymal Cells, J Cell Biol 43, 506, 1969
Collagenase Type 1: 0.05%

 

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