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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。
上皮细胞组织消化手册
| 物种 | 详细来源 | 细胞名称 | 消化用酶 | 培养基 | 参考文献 |
| 牛 | 牛, fetal | Epithelial | Neutral Protease: 2% | Dissociation medium, CMF | Schumann, B.L., Cody, T.E., Miller, M.L., Leikauf, G.D.: Isolation, Characterization, and Long-Term Culture of Fetal Bovine Tracheal Epithelial Cells, In Vitro Cell Dev Biol 24, 211, 1988 |
| Tracheal | |||||
| 犬 | 狗 | Tracheal | Pronase: 0.1% | DMEM | Virmani, A., Naziruddin, B., Desai, V., Lowry, J., Graves, D., and Sachdev, G.:Evidence for Secretion of High Molecular Weight Mucins by Canine Tracheal Epithelial Cells in Primary Culture: Effects of Select Secretagogues on Mucin Secretion, In Vitro Cell Dev Biol 28A, 120, 1992 |
| 鸡 | 小鸡, 5 天大 | Intestinal mesenchymal and epithelial | Collagenase: 0.03% | DMEM | Simon-Assmann, P and Kedinger, M: Embryonic Gut-Dissagregated Culture,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12A:3.1, 1995 |
| 鱼 | 鲨鱼(海绿角鲨) | Rectal gland | Collagenase: 0.2% | Ringer's solution | Karnaky, Jr., K.J., Valentich, J.D., Currie, M.G., Oehlenschlager, W.F., and Kennedy, M.P.: Atriopeptin Stimulates Chloride Secretion in Cultured Shark Rectal Gland Cells, Am J Physiol 260, 1125, 1991 |
| 鱼 | 鲨鱼(海绿角鲨) | Rectal gland | Collagenase: 0.2% | Ringer's solution | Valentich, J: J Tiss Cul Meth 13, 149, 1991 |
| 鱼 | 美洲拟鲽, 200-500 克 | Renal tubule | Trypsin: 0.2% | CMF solution | Dickman, K.G., and Renfro, J.: Primary Culture of Flounder Renal Tubule Cells: Transepithelial Transport, Am J Physiol 251, 424, 1986 |
| (黄盖美洲) | |||||
| 青蛙 | 青蛙, 有爪蟾蜍, 成年的, 雌性 | Colonic epithelial | Collagenase Type 4: 0.1% | Kreb's | Heinke, B, and Clauss, W.: Potassium Conductances in Isolated Single Cells fromXenopus Laevis Colonic Epithelium, J Comp Physiol [B] 169, 148, 1999 |
| 豚鼠 | Hartley豚鼠, 雌性, 200 克 | Epithelial | Collagenase Type 1: 0.1% | DMEM | Rutten, M.: Use of Commerically Available Cell Culture Inserts for Primary Culture and Electrophysiologic Studies of Guinea Pig Gastric Mucous Epithelial Cells, J Tiss Cul Meth 14, 235, 1992 |
| 豚鼠 | 豚鼠, 哈特利白化, | Endometrial | Collagenase: 0.25% | HBSS | Chaminadas, G., Alkhalaf, M., Remy-Martin, J.P., Propper, A.Y., and Adessi, G.L.: Specific Effect of Oestrone Sulphate on Protein Synthesis and Secretion by Cultured Epithelial Cells from Guinea-pig Endometrium, J Endocrinol 123, 233, 1989 |
| 500-600 克 | |||||
| 仓鼠 | 仓鼠, 叙利亚黄金, 雄性, 100-120 克 | Tracheal | Pronase: 0.1% | MEM with Hepes, CMF | Niles,R., Kim, K.C., Hyman, B., Christensen, T., Wasano, K., Brody, J.:Characterization Of Extended Primary And Secondary Cultures Of Hamster Tracheal Epithelial Cells, In Vitro Cell Dev Biol 24, 457, 1988 |
| 仓鼠 | 仓鼠, 叙利亚黄金 (铬：RGH应变) | Tracheal | Trypsin: 0.1% | Ham's F-12 | McDowell, E., et al.: Differentiation of Tracheal Mucociliary Epithelium in Primary Cell Culture Recapitulates Normal Fetal Development and Regeneration Following Injury in Hamsters, Am J Pathol 129, 511, 1987 |
| 仓鼠 | 仓鼠 (血统1516 EHS 和 莱克维尤), 8-12 周 | Tracheal | Trypsin: 0.05% | Medium 199 | Lee, T., Wu, R., Brody, A., Barrett, J., and Nettesheim, P.: Growth and Differentiation of Hamster Tracheal Epithelial Cells in Culture, Exp Lung Res 6, 27, 1984 |
| 仓鼠 | 仓鼠,叙利亚黄金, 雄性, 6 周- 4 个月 | Tracheal | Trypsin: 0.25% | PBS with EDTA | Goldman, W.E., Baseman, J.B.: Selective Isolation and Culture of a Proliferating Epithelial Cell Population from the Hamster Trachea, In Vitro 16, 313, 1980 |
| 人类 | 人类 | Human tracheal epithelium | Protease Type XIV: 0.04% | DMEM/F12 | Widdicombe JH, Sachs LA, Morrow JL, and Finkbeiner WE.: Expansion of cultures of human tracheal epithelium with maintenance of differentiated structure and function, Biotechniques 39(2), 249, 2005 |
| 人类 | 人类 | Colonic epithelial | Collagenase: | RPMI 1640 | Fukushima, K. and Fiocchi, C.: Paradoxical Decrease of Mitochondrial DNA Deletions in Epithelial Cells of Active Ulcerative Colitis Patients., Am J Physiol Gastrointest Liver Physiol 286, G804-13, 2004 |
| Neutral Protease: 0.3% | |||||
| Deoxyribonuclease I: 0.05% | |||||
| 人类 | 人类 | Corneal limbal epithelial sheet | Neutral Protease: 5% | see reference | Espana EM, Romano AC, Kawakita T, Di Pascuale M, Smiddy R, and Tseng SC:Novel enzymatic isolation of an entire viable human limbal epithelial sheet, Inv Ophthal Visual Sci 44(10), 4275, 2003 |
| 人类 | 人类 | Intestinal epithelial | Collagenase Type 4: 72.5 u/ml | HBSS | Fahlgren, A., Hammarstrom, S., Danielsson, A. and Hammarstrom, M.:Increased Expression of Antimicrobial Peptides and Lysozyme in Colonic Epithelial Cells of Patients with Ulcerative Colitis., Clin Exp Immunol 131, 90, 2003 |
| 人类 | 人类 | Gastric epithelial cells | Collagenase Type 2: 200 u/ml | L-15 | Smoot, D., Sewchand, J., Young, K., Desbrodes, B., Allen, C. and Naab, T.: A Method for Establishing Primary Cultures of Human Gastric Epithelial Cells,Meth Cell Sci 22, 133, 2000 |
| Neutral Protease: 1.2 u/ml | |||||
| Soybean Trypsin Inhibitor: 0.125% | |||||
| 人类 | 人类 | Gastric | Collagenase Type 4: 0.01% | F-12 medium | Sarosiek, J, Marshall, B, Peura, D, Guerrant, L, McCallum, R and Little, C: The Isolation and Maintenance of Human Gastric Epithelial Cells in Primary Culture,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12B:10.1, 1995 |
| 人类 | 人类 | Nasal polyp epithelial | Neutral Protease: .004% | see reference | Halbert CL, Alexander IE, Wolgamot GM, Miller AD: Adeno-associated virus vectors transduce primary cells much less efficiently than immortalized cells, J Virol 69, 1473-9, 1995 |
| Trypsin: 0.1% | |||||
| 人类 | 人类 | Epithelial | Trypsin: 0.05% | DMEM | Sabatini, L., Allen-Hoffmann, B, Warner, T., and Azen, E.: Serial C*tion of Epithelial Cells from Human and Macaque Salivary Glands, In Vitro Cell Dev Biol27A, 939, 1991 |
| 人类 | 人类 | Epithelial | Trypsin: 0.2% | MEM, PBS | Robinson, C., and Wu, R.: Culture of Conducting Airway Epithelial Cells in Serum-Free Medium, J Tiss Cul Meth 13, 95, 1991 |
| 人类 | 人类 | Epithelial | Collagenase: 2.0% | DMEM/Ham's F-12 | Emerman, J. and Wilkinson, D.: Routine Culturing of Normal, Dysplastic and Malignant Human Mammary Epithelial Cells from Small Tissue Samples, In Vitro Cell Dev Biol 26, 1186, 1990 |
| 人类 | 人类 | Epithelial | Pronase: 0.1% | PBS | Gruenert, D.C., Basbaum, C.B., and Widdicombe, J.H.: Long-Term Culture of Normal and Cystic Fibrosis Epithelial Cells Grown Under Serum-Free Conditions, In Vitro Cell Dev Biol 26, 411, 1990 |
| 人类 | 人类 | Epithelial | Collagenase Type 2: 0.2% | (see reference) | Wood, L. and Neufeld, E.: A Cystic Fibrosis Phenotype in Cells Cultured from Sweat Gland Secretory Coil. Altered Kinetics of 36Cl Efflux, J Biol Chem 265, 12796, 1990 |
| Sweat gland | |||||
| 人类 | 人类 | Endometrial epithelial | Trypsin: | DMEM/Ham's F-12 | Munson, L., Chandler, S., and Schlafer, D.: C*tion of Bovine Fetal and Adult Endometrial Epithelial Cells, J Tiss Cul Meth 11 (3), 129, 1988 |
| (包括 牛) | |||||
| 人类 | 人类 | Epithelial | Deoxyribonuclease I: 0.01% | HEPES with 5.9mM Glucose,5mM DTT | Widdicombe, J.H., Coleman, D.L., Finkbeiner, W.E., and Tuet, I.K.: Electrical Properties of Monolayers Cultured From Cells of Human Tracheal Mucosa, J Appl Physiol 58, 1729, 1985 |
| 人类 | 人类 | Epithelial | Protease Type XIV: 0.1% | Eagle's MEM | Yankaskas, J., Cotton, C., Knowles, M., Gatzy, J., and Boucher, R.: Culture of Human Nasal Epithelial Cells on Collagen Matrix Supports, Am Rev Respir Dis 132, 1281, 1985 |
| 人类 | 人类,女人, 27-49 岁 | Epithelial | Trypsin: 0.125% | HBSS, CMF | Auersperg, N., Siemens, C.H., and Myrdal, S.E.: Human Ovarian Surface Epithelium In Primary Culture, In Vitro 20, 743, 1984 |
| Ovary | |||||
| 人类 | 人类,婴儿和新生儿 | Epithelial | Trypsin: 0.1% | HBSS | Lechner, J., Babcock, M., Marnell, M., Narayan, K., and Kaighn, M.: Normal Human Prostate Epithelial Cell Cultures, Methods Cell Biol 21, 195, 1980 |
| Prostate | |||||
| 小鼠 | 小鼠, 成年的 | Cilary epithelial | PDS kit: per instructions | EBSS | Gualdoni, S., Baron, M., Lakowski, J., Decembrini, S., Pearson, R., Ali, R. and Sowden, J.: Isolation and Culture of Adult Ciliary Epithelial Cells, Previously Identified as Retinal Stem Cells, and Retinal Progenitor Cells., Curr Protoc Stem Cell Biol Chapter 1, Unit 1H.4, 2011 |
| 小鼠 | 小鼠, 雄性, 8-16 周 | Renal tubular epithelial | Collagenase: 200 u/ml | HBSS | Breggia, A. and Himmelfarb, J.: Primary Mouse Renal Tubular Epithelial Cells have Variable Injury Tolerance to Ischemic and Chemical Mediators of Oxidative Stress.,Oxid Med Cell Longev 1, 33, 2008 |
| Soybean Trypsin Inhibitor: see reference | |||||
| 小鼠 | 小鼠, 6-8 周 | Lamina propria mononuclear cells | Collagenase Type 2: 0.015% | RPMI | Wirtz S., Becker C., Blumberg R., Galle P., and Neurath M.: Treatment of T cell-dependent experimental colitis in SCID mice by local administration of an adenovirus expressing IL-18 antisense mRNA, J Immunol 168(1), 411, 2002 |
| Deoxyribonuclease I: 0.01% | |||||
| 小鼠 | 小鼠, 11 周 | Epithelial | Collagenase Type 3: 25 u/ml | DMEM/F12 | Mueller, S., Clark, J., Myers, P. and Korach, K.: Mammary Gland Development in Adult Mice Requires Epithelial and Stromal Estrogen Receptor Alpha.,Endocrinology 143, 2357, 2002 |
| Hyaluronidase: 0.1% | |||||
| Protease XIV: 0.05% | |||||
| Deoxyribonuclease I: 0.04% | |||||
| 小鼠 | 小鼠, 雌性 | Salivary gland epithelial | Collagenase Type 1: 750 u/ml | DMEM/F12 | Ishimaru N, Saegusa K, Yanagi K, Haneji N, Saito I, Hayashi Y: Estrogen deficiency accelerates autoimmune exocrinopathy in murine Sjogren's syndrome through fas-mediated apoptosis, Am J Pathol 155, 173-81, 1999 |
| Hyaluronidase: 500 u/ml | |||||
| 小鼠 | 小鼠, 胎儿12-13 天 | Intestinal mesenchymal and epithelial | Collagenase: 0.03% | DMEM | Simon-Assmann, P and Kedinger, M: Embryonic Gut-Dissagregated Culture,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12A:3.1, 1995 |
| 小鼠 | 小鼠 | Submandibular salivary | Collagenase Type 2 or 3: 0.16% | DMEM | Durban, E: Submandibular Salivary Epithelial Cells,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12B:2.1, 1995 |
| 小鼠 | 小鼠 | Esophageal | Trypsin: 0.25% | PBS, CMF | Katayama, M., Kan, M.: Heparin-Binding (Fibroblast) Growth Factors re Potential Autocrine Regulators of Esophageal Epithelial Cell Proliferation, In Vitro Cell Dev Biol 27, 533, 1991 |
| 小鼠 | 小鼠, 成年的, 雌性, 6-8周大 | Uterine | Trypsin: 0.2% | HBSS | Ghosh, D., Danielson, K., Alston, J., and Heyner, S.: Functional Differentiation of Mouse Uterine Epithelial Cells Grown on Collagen Gels or Reconstituted Basement Membranes, In Vitro Cell Dev Biol 27A, 713, 1991 |
| 小鼠 | 小鼠, 雌性, 18 - 20天 | Uterine | Trypsin: 0.25% | HBSS | Fukamachi, H., and McLachlan, J.: Proliferation and Differentiation of Mouse Uterine Epithelial Cells in Primary Serum-Free Culture: Estradiol-17B Suppresses Uterine Epithelial Proliferation Cultured on a Basement Membrane-Like Substratum,In Vitro Cell Dev Biol 27A, 907, 1991 |
| (包括 20 - 22 天) | |||||
| 小鼠 | 小鼠, (白变种实验室老鼠), 雄性, 3-5 月 | Epithelial | Collagenase Type 3: 0.16% , 1:1 v/v | DMEM with 15 mM HEPES | Durban, E.M.: Mouse Submandibular Salivary Epithelial Cell Growth and Differentiation in Long-Term Culture:Influence of the Extracellular Matrix, In Vitro Cell Dev Biol 26, 33, 1990 |
| Submandibular salivary gland | |||||
| 小鼠 | 小鼠, 雌性 | Epithelial | Pepsin: 0.1% | HBSS | Reiser, M., Huff, B., and Medina, D.: Pepsin Can be Used to Subculture Viable Mammary Epithelial Cells, In Vitro 19 (9), 730, 1983 |
| 小鼠 | 小鼠, (白变种实验室老鼠fC3H or白变种实验室老鼠) 8-12 天中孕 | Epithelial | Collagenase Type 3: 0.1% | HBSS | Yang, J., Flynn, D., Larson, L., and Hamamoto, S.: Growth in Primary Culture of Mouse Submandibular Epithelial Cells, In Vitro 18, 435, 1982 |
| Submandibular gland | |||||
| 小鼠 | 小鼠 (白变种实验室老鼠fC3H) | Mammary tumors | Collagenase: 1.0% | HBSS | Yang, J., Guzman, R., Richards, J., and Nandi, S.: Primary Cultures of Mouse Mammary Tumor Epithelial Cells Embedded in Collagen Gels, In Vitro 16, 502, 1980 |
| Epithelial | |||||
| 小鼠 | 小鼠, C3H, 6-8 周 | 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 |
| 猪 | 猪, 5-60 公斤 | Retinal pigment epithelial cells | Collagenase: 2% | DMEM | Wiencke, A., Kiilgaard, J., Nicolini, J., Bundgaard, M., Ropke, C., and La Cour, M.: Growth of Cultured Porcine Retinal Pigment Epithelial Cells, Acta Opthalmol Scand 81(2), 170, 2003 |
| 猪 | 猪 | Trachea | Neutral Protease: 0.2% | HBSS | De Buysscher, E., Kennedy, J., and Mendicino, J.: Synthesis of Mucin Glycoproteins by Epithelial Cells Isolated from Swine Trachea by Specific Proteolysis, In Vitro 20, 433, 1984 |
| 兔 | 兔, 新西兰白, 成年的 | Colon | Neutral Protease: 0.3% | PBS | Vidrich, A., Racindranath, R., Farsi, K., and Targan, S.: A Method for the Rapid Establishment of Normal Adult Mammalian Colonic Epithelial Cell Cultures, In Vitro Cell Dev Biol 24 (3), 188, 1988 |
| 兔 | 兔, 新西兰白, 雄性, 4-5磅 | Gastric | Collagenase Type 2: 0.08% | Sodium phosphate buffer | Chew, C.S., Brown,M.R.: Release of Intracellular Ca2+ and Elevation of Inositol Triphosphate by Secretagogues in Parietal and Chief Cells Isolated from Rabbit Gastric Mucosa, Biochim Biophys Acta 888, 116, 1986 |
| Parietal and chief | |||||
| 兔 | 兔, 新西兰白 动情期的,雌性, 4-5 个月 | Mesothelial and surface epithelial | Trypsin: 0.125%-0.5% | Medium 199 | Nicosia, S., Johnson, J., and Streibel, E.: Isolation and Ultrastructure of Rabbit Ovarian Mesothelium(Surface Epithelium), Int J Gynecol Pathol 3, 348, 1984 |
| Ovaries | |||||
| 兔 | 兔, 胎儿的 | Epithelial | Collagenase Type 3: 0.10% | HBSS | Logsdon, C.D., Bisbee, C.A., Rutten, M.J. and Machen, T.E.: Fetal Rabbit Gastric Epithelial Cells Cultured on Floating Collagen Gel, In Vitro 18, 233, 1981 |
| Gastric | |||||
| 大鼠 | 大鼠 | Mammary epithelial | Collagenase Type 3: 0.35% | HBSS | Mei, N., McDaniel, L., Dobrovolsky, V., Guo, X., Shaddock, J., Mitstaedt, R., Azuma, M., Shelton, S., McGarrity, L., Doerge, D. and Heflich, R.: The Genotoxicity of Acrylamide and Glycidamide in Big Blue Rats., Toxicol Sci 115, 412, 2010 |
| 大鼠 | 大鼠, SD, 成年的,男性, 8-10 周大 | Seminiferous tubules | Trypsin: 0.05% | Krebs-Ringer bicarbonate buffer | Abou-Haila, A., and Tulsiani, D.: Acid Glycohydrolases in Rat Spermatocytes, Spermatids and Spermatozoa: Enzyme Activities, Biosynthesis and Immunolocalization, Biol Proced Online 3 (1), 35, 2001 |
| (see reference) | |||||
| 大鼠 | 大鼠, 男性 | Alveolar epithelial | Elastase: 40 u/ml | DMEM | Planus, E., Galiacy, S., Matthay, M., Laurent, V., Gavrilovic, J., Murphy, G., Clerici, C., Isabey, D., Lafuma, C., and d'Ortho, M.: Role of Collagenase in Mediating in Vitro Alveolar Epithelial Wound Repair, J Cell Sci 112, 243, 1999 |
| 大鼠 | 大鼠, 胚胎 | Tracheal epithelial | Collagenase Type 4: 0.05% | DMEM/F12 | Shannon JM, Gebb SA, and Nielsen LD: Induction of alveolar type II cell differentiation in embryonic tracheal epithelium in mesenchyme-free culture,Development 126, 1675, 1999 |
| Neutral Protease: | |||||
| Deoxyribonuclease I: | |||||
| 大鼠 | 大鼠, fetal, 14-15 day | Intestinal mesenchymal and epithelial | Collagenase: 0.03% | DMEM | Simon-Assmann, P and Kedinger, M: Embryonic Gut-Dissagregated Culture,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 12A:3.1, 1995 |
| 大鼠 | 大鼠, 4-14 周 | Retinal pigment epithelial cells | Collagenase Type 1: 65 u/ml | CF Hanks with EDTA | Wang N, Koutz CA, and Anderson RE.: A method for the isolation of retinal pigment epithelial cells from adult rats, Inv Ophthal Visual Sci 34(1), 101, 1993 |
| Hyaluronidase: 220 u/ml | |||||
| 大鼠 | 大鼠, SD, 成年的, 90-120 日大, 350-450克 | Epididymal epithelial | Collagenase Type 2: 0.1% | HBSS | Klinefelter, G.: A Novel System for the Co-Culture of Epididymal Epithelial Cells and Sperm From Adult Rats, J Tiss Cul Meth 14, 195, 1992 |
| 大鼠 | 大鼠, 6 天大 | Rat intestinal epithelial | Neutral Protease: 0.01% | DMEM | Evans GS, Flint N, Somers AS, Eyden B, and Potten CS.: The development of a method for the preparation of rat intestinal epithelial cell primary cultures, J Cell Sci 101, 219, 1992 |
| Collagenase: 300 u/ml | |||||
| 大鼠 | 大鼠, SD, 雄性, 150 - 250克 | Epithelial | Pronase: 0.15% | Medium 199 | Dial, E., Kao, Y., and Lichtenberger, L.: Effects of 16,16-Dimethyl Prostaglandin E2 On Glycoprotein And Lipid Synthesis of Gastric Epithelial Cells Grown in a Primary Culture, In Vitro Cell Dev Biol 27, 39, 1991 |
| Stomach | |||||
| 大鼠 | 大鼠, 成年的 | Interlobular duct fragments | Papain: | DMEM/Ham's F-12 | Heimann, T., and Githens, S.: Rat Pancreatic Duct Epithelium Cultured on a Porous Support Coated with Extracellular Matrix, Pancreas 6 (5), 514, 1991 |
| (包括 仓鼠) | |||||
| 大鼠 | 大鼠, SD, 200 克 | Colon | Deoxyribonuclease I: 10 礸/ml | (see reference) | Yassin, R., Clearfield, H., Katz, S., and Murthy, S.: Gastrin Induction of mRNA Expression in Rat Colonic Epithelium In Vitro, Peptides 12, 63, 1991 |
| 大鼠 | Wistar大鼠, 新生的 | Epithelial | Trypsin: 0.1% | HBSS | Jassal, D., Han, R., Caniggia, I., Post, M., and Tanswell, A.: Growth of Distal Fetal Rat Lung Epithelial Cells in a Defined Serum-Free Medium, In Vitro Cell Dev Biol27A, 625, 1991 |
| 大鼠 | 啮齿目动物, 各种各样的 (参考) | Epithelial | Trypsin: 0.2% | MEM, PBS | Robinson, C., and Wu, R.: Culture of Conducting Airway Epithelial Cells in Serum-Free Medium, J Tiss Cul Meth 13, 95, 1991 |
| 大鼠 | 大鼠, 6-8 天 | Retinal pigment epithelial | Neutral Protease: 2% | DMEM | Chang CW, Roque RS, Defoe DM, and Caldwell RB.: An improved method for isolation and culture of pigment epithelial cells from rat retina, Curr Eye Res10(11), 1081, 1991 |
| 大鼠 | 大鼠 (ACI/NMs X BUF/Mna) F1, 雄性, 28个 月 | Epthelial | Collagenase Type 3: 0.1% | Eagle's MEM Serum-free | Masuda, A., Ohtsuka, K., and Matsuyama, M.: Establishment of Functional Epithelial Cell Lines from a Rat Thyoma and a Rat Thymus, In Vitro Cell Dev Biol 26, 713, 1990 |
| 大鼠 (ACI/MNs) 雄性, 8周 | |||||
| 大鼠 | 大鼠, 费希尔344, 雄性, 8 周大, | Tracheal epithelial | Pronase: 0.5% | DMEM | Chang, L., Wu, R., and Nettesheim, P.: Morphological Changes in Rat Tracheal Cells During The Adaptive and Early Growth Phase in Primary Cell Culture, J Cell Sci74, 283, 1985 |
| 250-300 克 | |||||
| 大鼠 | 大鼠,费希尔, 雄性, 4-6 周 | Epithelial | Hyaluronidase: 0.1% | HEPES BSS | Babcock, M., Marino, M., Gunning, W., and Stoner, G.: Clonal Growth and Serial Propagation of Rat Esophageal Epithelial Cells, In Vitro 19, 403, 1983 |
| Esophagus | |||||
| 大鼠 | 大鼠, 费希尔, Lewis and SD, 雄性, 10-18 个月 | 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 | 大鼠, SD, 雌性 | Epithelial, cancer and tumor | Collagenase: 0.1% | Eagles's MEM | Cohen, L.: Isolation and Characterization of a Serially C*ted, Neoplastic, Epithelial Cell Line from the N-nitrosomethylurea Induced Rat Mammary Adenocarcinoma, In Vitro 18, 565, 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 | 大鼠,费希尔, 成年的, 200-250 克 | Epithelial | Hyaluronidase: 0.0075% | KCl-NaCl HEPES Buffer | Williams, G., and Gunn, J.: Long-Term Culture of Adult Rat Liver Epithelial Cells,Exp Cell Res 89, 139, 1974 |
| 大鼠 | Fischer大鼠, 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 |
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