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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(méi)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(guò)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(lái)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(méi)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(guò)查閱文獻(xiàn)來(lái)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(lái)源于組織胞外基質(zhì)的復(fù)雜而動(dòng)態(tài)的特性，以及過(guò)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， 以便于制定一個(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(jiàn)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 |
眼 | 心臟 | 腸 | 腎 |
肝 | 肺 | 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。
上皮細(xì)胞組織消化手冊(cè)
| 物種 | 詳細(xì)來(lái)源 | 細(xì)胞名稱 | 消化用酶 | 培養(yǎng)基 | 參考文獻(xiàn) |
| 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 |
| 雞 | 小雞, 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 |
| 魚(yú) | 鯊魚(yú)(海綠角鯊) | 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 |
| 魚(yú) | 鯊魚(yú)(海綠角鯊) | Rectal gland | Collagenase: 0.2% | Ringer's solution | Valentich, J: J Tiss Cul Meth 13, 149, 1991 |
| 魚(yú) | 美洲擬鰈, 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 |
| (黃蓋美洲) | |||||
| 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 | |||||
| 倉(cāng)鼠 | 倉(cāng)鼠, 敘利亞黃金, 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 |
| 倉(cāng)鼠 | 倉(cāng)鼠, 敘利亞黃金 (鉻：RGH應(yīng)變) | 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 |
| 倉(cāng)鼠 | 倉(cāng)鼠 (血統(tǒng)1516 EHS 和 萊克維尤), 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 |
| 倉(cāng)鼠 | 倉(cāng)鼠,敘利亞黃金, 雄性, 6 周- 4 個(gè)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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 | |||||
| 人類 | 人類 | 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) | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類,女人, 27-49 歲 | 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 | |||||
| 人類 | 人類,嬰兒和新生兒 | 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, 胎兒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, (白變種實(shí)驗(yàn)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, (白變種實(shí)驗(yàn)室老鼠fC3H or白變種實(shí)驗(yàn)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 (白變種實(shí)驗(yàn)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 |
| 豬 | 豬, 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 |
| 豬 | 豬 | 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, 新西蘭白, 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, 新西蘭白, 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, 新西蘭白 動(dòng)情期的,雌性, 4-5 個(gè)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, 胎兒的 | 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 倉(cāng)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 | 嚙齒目動(dòng)物, 各種各樣的 (參考) | 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個(gè) 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, 費(fèi)希爾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,費(fèi)希爾, 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, 費(fèi)希爾, Lewis and SD, 雄性, 10-18 個(gè)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,費(fèi)希爾, 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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