worthington 上皮細胞組織分離指南

發(fā)布者：艾美捷科技發(fā)布時間：2022-06-01

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原代細胞分離中，針對上皮細胞組織涉及到上皮，氣管、直腸腺、結腸上皮和胃腸上皮、鼻上皮等等的解離通常會使用酶，如何選擇合適的酶來完成對應上皮細胞組織的解離，以及原代細胞采集的常規(guī)操作， 艾美捷針對上皮細胞相關的組織分類解離分種屬、不同的細胞、酶組合推薦，推薦濃度、培養(yǎng)基以及參考文獻，詳細匯總指南：

上皮組織分離指南

本技術指南描述了標準實驗室程序；為建立細胞分離方案提供了一種合乎邏輯的實驗方法；并列出了許多組織特異性參考資料，以幫助開發(fā)有效的方法。更多其他組織的解離指南，點擊查看：各組織分離指南。

上皮細胞
種屬	種屬詳情	細胞名稱	酶組合推薦，含推薦濃度	培養(yǎng)基	參考文獻
牛	Bovine, fetal	上皮，氣管 / Epithelial，Tracheal	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
犬/狗	Dog	氣管 / 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	Chick, 5 day old	腸間充質和上皮 / 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	Shark (Squalus acanthias)	直腸腺 / 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	Shark (Squalus acanthias)	直腸腺 / Rectal gland	Collagenase: 0.2%	Ringer's solution	Valentich, J: Primary Cultures of Shark Rectal Gland Epithelial Cells: A Model for Hormone-Sensitive Chloride Transport, J Tiss Cul Meth 13, 149, 1991
魚	Winter flounder, 200-500 g	腎小管 / Renal tubule	Trypsin: 0.2%	CMF solution	Dickman, K. and Renfro, J.: Primary Culture of Flounder Renal Tubule Cells:Transepithelial Transport, Am J Physiol 251, 424, 1986
青蛙	Frog, Xenopus laevis, adult, female	結腸上皮 / Colonic epithelial	Collagenase Type 4: 0.1%	Kreb's	Heinke, B, and Clauss, W.: Potassium Conductances in Isolated Single Cells from Xenopus Laevis Colonic Epithelium, J Comp Physiol [B] 169, 148, 1999
豚鼠	Guinea-pig, Hartley, female, 200 g	上皮 / 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	Guinea-pig, Hartley albino,	子宮內膜 / 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
倉鼠	Hamster, Syrian gold, male, 100-120 g	氣管 / 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
倉鼠	Hamster, Syrian golden (strain CR:RGH)	氣管 / 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
倉鼠	Hamster (strain 1516 EHS and Lakeview), 8-12 weeks	氣管 / 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
倉鼠	Hamster, Syrian gold, male, 6 weeks- 4 months	氣管 / 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	Human	上皮 / Epithelial	Collagenase Type 4: 0.1%； Neutral Protease: 0.1%	PBS	Gottipamula, S., Saraswat, S. and Sridhar, K.: Comparative Study of Isolation, Expansion and Characterization of Epithelial Cells., Cytotherapy 19, 263-271, 2017
人	Human	口腔黏膜上皮 / Oral mucosal epithelial	Collagenase Type 1: 0.05%	DMEM/F12	Hsueh, Y., Huang, S., Lai, J., Ma, S., Chen, H., Wu, S., Wang, T., Sun, C., Ma, K., Chen, J., Lai, C. and Ma, D.: Preservation of Epithelial Progenitor Cells from Collagenase-Digested Oral Mucosa During Ex Vivo Cultivation., Sci Rep 6, 36266, 2016
人	Human	胃腸上皮 / Gastrointestinal epithelial	Collagenase Type 1: 0.2%	DMEM/F12	VanDussen, K., Marinshaw, J., Shaikh, N., Miyoshi, H., Moon, C., Tarr, P., Ciorba, M. and Stappenbeck, T.: Development of an Enhanced Human Gastrointestinal Epithelial Culture System to Facilitate Patient-Based Assays., Gut 64, 911-20, 2015
人	Human	人氣管上皮 / 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	Human	結腸上皮 / Colonic epithelial	Collagenase:； Neutral Protease: 0.3%； Deoxyribonuclease I: 0.05%	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
人	Human	角膜緣上皮片 / 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	Human	腸上皮 / 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	Human	胃上皮細胞 / Gastric epithelial cells	Collagenase Type 2: 200 u/ml； Neutral Protease: 1.2 u/ml； Soybean Trypsin Inhibitor: 0.125%	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
人	Human	胃 / 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	Human	鼻息肉上皮 / Nasal polyp epithelial	Neutral Protease: .004%； Trypsin: 0.1%	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
人	Human	上皮 / Epithelial	Trypsin: 0.05%	DMEM	Sabatini, L., Allen-Hoffmann, B, Warner, T., and Azen, E.: Serial Cultivation of Epithelial Cells from Human and Macaque Salivary Glands, In Vitro Cell Dev Biol 27A, 939, 1991
人	Human	上皮 / 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	Human	上皮 / 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	Human	上皮 / 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	Human	上皮 / 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
人	Human	子宮內膜上皮 / Endometrial epithelial	Trypsin:	DMEM/Ham's F-12	Munson, L., Chandler, S., and Schlafer, D.: Cultivation of Bovine Fetal and Adult Endometrial Epithelial Cells, J Tiss Cul Meth 11 (3), 129, 1988
人	Human	上皮 / 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	Human	上皮 / 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	Human, women, 27-49 years	上皮 / 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
人	Human, infant and neonate	上皮 / 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
小鼠	Mouse, 8-12 week	鼻上皮 / Nasal epithelium	Papain: 20 u/ml； Deoxyribonuclease I: 0.005%	EBSS	Brann, D., Tsukahara, T., Weinreb, C., Lipovsek, M., Van den Berge, K., Gong, B., Chance, R., Macaulay, I., Chou, H., Fletcher, R., Das, D., Street, K., de Bezieux, H., Choi, Y., Risso, D. and Dudoit, S.: Non-Neuronal Expression of SARS-CoV-2 Entry Genes in the Olfactory System Suggests Mechanisms Underlying COVID-19-Associated Anosmia., Sci. Adv. 6, , 2020
小鼠	Mouse	肺泡上皮 / Alveolar epithelial	Neutral Protease: 0.1%； Deoxyribonuclease I: 0.01%	DMEM	Sun, F., Xiao, G. and Qu, Z.: Isolation of Murine Alveolar Type II Epithelial Cells., Bio-protocol 7, , 2017
小鼠	Mouse	氣管上皮 / Tracheal epithelial	Papain: 50 u/ml	See reference	Pardo-Saganta, A., Tata, P., Law, B., Saez, B., Chow, R., Prabhu, M., Gridley, T. and Rajagopal, J.: Parent Stem Cells Can Serve as Niches for Their Daughter Cells., Nature 523, 597-601, 2015