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worthington 上皮细胞组织分离指南

发布者:艾美捷科技    发布时间:2022-06-01     
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原代细胞分离中,针对上皮细胞组织涉及到上皮,气管、直肠腺、结肠上皮和胃肠上皮、鼻上皮 等等的解离通常会使用酶,如何选择合适的酶来完成对应上皮细胞组织的解离,以及原代细胞采集的常规操作, 艾美捷针对上皮细胞相关的组织分类解离分种属、不同的细胞、酶组合推荐,推荐浓度、培养基以及参考文献,详细汇总指南:


上皮组织分离指南

本技术指南描述了标准实验室程序; 为建立细胞分离方案提供了一种合乎逻辑的实验方法; 并列出了许多组织特异性参考资料,以帮助开发有效的方法。更多其他组织的解离指南,点击查看:各组织分离指南

上皮细胞
种属种属详情细胞名称酶组合推荐,含推荐浓度培养基参考文献
Bovine, fetal上皮,气管 / Epithelial,TrachealNeutral Protease: 2%Dissociation medium, CMFSchumann, 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气管 / TrachealPronase: 0.1%DMEMVirmani, 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
Chick, 5 day old肠间充质和上皮 / Intestinal mesenchymal and epithelialCollagenase: 0.03%DMEMSimon-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
Shark (Squalus acanthias)直肠腺 / Rectal glandCollagenase: 0.2%Ringer's solutionKarnaky, 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
Shark (Squalus acanthias)直肠腺 / Rectal glandCollagenase: 0.2%Ringer's solutionValentich, J: Primary Cultures of Shark Rectal Gland Epithelial Cells: A Model for Hormone-Sensitive Chloride Transport, J Tiss Cul Meth 13, 149, 1991
Winter flounder, 200-500 g肾小管 / Renal tubuleTrypsin: 0.2%CMF solutionDickman, 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结肠上皮 / Colonic epithelialCollagenase Type 4: 0.1%Kreb'sHeinke, 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 / EpithelialCollagenase Type 1: 0.1%DMEMRutten, 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,子宫内膜 / EndometrialCollagenase: 0.25%HBSSChaminadas, 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
仓鼠Hamster, Syrian gold, male, 100-120 g气管 / TrachealPronase: 0.1%MEM with Hepes, CMFNiles,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Hamster, Syrian golden (strain CR:RGH)气管 / TrachealTrypsin: 0.1%Ham's F-12McDowell, 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Hamster (strain 1516 EHS and Lakeview), 8-12 weeks气管 / TrachealTrypsin: 0.05%Medium 199Lee, 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Hamster, Syrian gold, male, 6 weeks- 4 months气管 / TrachealTrypsin: 0.25%PBS with EDTAGoldman, W.E., Baseman, J.B.: Selective Isolation and Culture of a Proliferating Epithelial Cell Population from the Hamster Trachea, In Vitro 16, 313, 1980
Human上皮 / EpithelialCollagenase Type 4: 0.1%;
Neutral Protease: 0.1%
PBSGottipamula, S., Saraswat, S. and Sridhar, K.: Comparative Study of Isolation, Expansion and Characterization of Epithelial Cells., Cytotherapy 19, 263-271, 2017
Human口腔黏膜上皮 / Oral mucosal epithelialCollagenase Type 1: 0.05%DMEM/F12Hsueh, 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
Human胃肠上皮 / Gastrointestinal epithelialCollagenase Type 1: 0.2%DMEM/F12VanDussen, 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
Human人气管上皮 / Human tracheal epitheliumProtease Type XIV: 0.04%DMEM/F12Widdicombe 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
Human结肠上皮 / Colonic epithelialCollagenase:;
Neutral Protease: 0.3%;
Deoxyribonuclease I: 0.05%
RPMI 1640Fukushima, 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
Human角膜缘上皮片 / Corneal limbal epithelial sheetNeutral Protease: 5%see referenceEspana 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
Human肠上皮 / Intestinal epithelialCollagenase Type 4: 72.5 u/mlHBSSFahlgren, 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
Human胃上皮细胞 / Gastric epithelial cellsCollagenase Type 2: 200 u/ml;
Neutral Protease: 1.2 u/ml;
Soybean Trypsin Inhibitor: 0.125%
L-15Smoot, 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
Human胃 / GastricCollagenase Type 4: 0.01%F-12 mediumSarosiek, 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
Human鼻息肉上皮 / Nasal polyp epithelialNeutral Protease: .004%;
Trypsin: 0.1%
see referenceHalbert 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
Human上皮 / EpithelialTrypsin: 0.05%DMEMSabatini, 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
Human上皮 / EpithelialTrypsin: 0.2%MEM, PBSRobinson, C., and Wu, R.: Culture of Conducting Airway Epithelial Cells in Serum-Free Medium, J Tiss Cul Meth 13, 95, 1991
Human上皮 / EpithelialCollagenase: 2.0%DMEM/Ham's F-12Emerman, 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
Human上皮 / EpithelialPronase: 0.1%PBSGruenert, 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
Human上皮 / EpithelialCollagenase 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
Human子宫内膜上皮 / Endometrial epithelialTrypsin:DMEM/Ham's F-12Munson, L., Chandler, S., and Schlafer, D.: Cultivation of Bovine Fetal and Adult Endometrial Epithelial Cells, J Tiss Cul Meth 11 (3), 129, 1988
Human上皮 / EpithelialDeoxyribonuclease I: 0.01%HEPES with 5.9mM Glucose,5mM DTTWiddicombe, 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
Human上皮 / EpithelialProtease Type XIV: 0.1%Eagle's MEMYankaskas, 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
Human, women, 27-49 years上皮 / EpithelialTrypsin: 0.125%HBSS, CMFAuersperg, N., Siemens, C.H., and Myrdal, S.E.: Human Ovarian Surface Epithelium In Primary Culture, In Vitro 20, 743, 1984
Human, infant and neonate上皮 / EpithelialTrypsin: 0.1%HBSSLechner, 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 epitheliumPapain: 20 u/ml;
Deoxyribonuclease I: 0.005%
EBSSBrann, 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 epithelialNeutral Protease: 0.1%;
Deoxyribonuclease I: 0.01%
DMEMSun, F., Xiao, G. and Qu, Z.: Isolation of Murine Alveolar Type II Epithelial Cells., Bio-protocol 7, , 2017
小鼠Mouse气管上皮 / Tracheal epithelialPapain: 50 u/mlSee referencePardo-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


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