worthington 皮肤组织分离指南

发布者：艾美捷科技发布时间：2022-06-09

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原代细胞分离中，针对皮肤组织涉及到表皮，真皮分离的聚集蛋白聚糖敏感、二倍体成纤维细胞、皮肤成纤维细胞，皮肤和角质形成细胞等等的解离通常会使用酶，如何选择合适的酶来完成对应皮肤组织的解离，以及原代细胞采集的常规操作， 艾美捷针对皮肤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：皮肤组织分离指南.png

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。

皮肤
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
犬/狗	Canine	表皮 / Epidermal	Collagenase: 0.35%； Neutral Protease: 0.1%	DMEM	Zheng, Y., Nace, A., Chen, W., Watkins, K., Sergott, L., Homan, Y., Vandeberg, J., Breen, M. and Stenn, K.: Mature Hair Follicles Generated from Dissociated Cells: A Universal Mechanism of Folliculoneogenesis., Dev Dyn 239, 2619-26, 2010
青蛙	Frog	表皮 / Epidermal	Trypsin: 0.18%	Barth's solution, CMF	Nishikawa, A., Shimizu-Nishikawa, K., and Miler, L.: Isolation, characterization, and in vitro culture of larval and adult epidermal cells of the frog Xenopus laevis, In Vitro Cell Dev Biol 26, 1128, 1990
山羊	Goat	真皮分离的聚集蛋白聚糖敏感 / Dermis isolated aggrecan sensitive	Neutral Protease: 0.5%； Collagenase Type 2: 200 u/ml	DMEM	Deng, Y., Hu, J. and Athanasiou, K.: Isolation and Chondroinduction of a Dermis-Isolated, Aggrecan-Sensitive Subpopulation with High Chondrogenic Potential., Arthritis Rheum 56, 168, 2007
人	Human	二倍体成纤维细胞 / Diploid fibroblasts	Collagenase Type 1: 0.03%	DMEM	Tan, J., Jaafar, F. and Makpol, S.: Proteomic Profiling of Senescent Human Diploid Fibroblasts Treated with gamma-tocotrienol., BMC Complement Altern Med 18, 314, 2018
人	Human	成纤维细胞 / Fibroblasts	Neutral Protease: 0.1%； Collagenase Type 1: 0.25%	DMEM	Chen, Z., Wang, X., Jin, T., Wang, Y., Hong, C., Tan, L., Dai, T., Wu, L., Zhuang, Z. and Shi, C.: Increase in the Radioresistance of Normal Skin Fibroblasts but not Tumor Cells by Mechanical Injury., Cell Death Dis 8, e2573, 2017
人	Human	单核吞噬细胞 / Mononuclear phagocytes	Neutral Protease: 1 u/ml； Deoxyribonuclease I: 50 u/ml； Collagenase: 200 u/ml； Trypsin: 0.5%；	RPMI-1640	Botting, Ra., Bertram, K., Baharlou, H., Sandgren, K., Fletcher, J., Rhodes, J., Rana, H., Plasto, T., Wang, X., Lim, J., Barnouti, L., Kohout, M., Papadopoulos, T., Merten, S., Olbourne, N. and Cunningham, A.: Phenotypic and Functional Consequences of Different Isolation Protocols on Skin Mononuclear Phagocytes., J Leukoc Biol 101, 1393-1403, 2017
人	Human	皮肤 / Skin	Neutral Protease: 5 u/ml； Collagenase Type 3: 0.3%； Deoxyribonuclease I: 0.005%；	RPMI 1640	Cheuk, S., Wiken, M., Blomqvist, L., Nylen, S., Talme, T., Stahle, M.and Eidsmo, L.: Epidermal Th22 and Tc17 Cells Form a Localized Disease Memory in Clinically Healed Psoriasis., J Immunol 192, 3111, 2014
人	Human	皮肤成纤维细胞 / Skin fibroblasts	Collagenase Type 1: 1000 u/ml	DMEM	Douvaras, P., Wang, J., Zimmer, M., Hanchuk, S., O'Bara, M., Sadiq, S., Sim, F., Goldman, J. and Fossati, V.: Efficient Generation of Myelinating Oligodendrocytes from Primary Progressive Multiple Sclerosis Patients by Induced Pluripotent Stem Cells., Stem Cell Reports 3, 250, 2014
人	Human	皮肤 / Skin	Collagenase Type 4: 0.08-0.2%； Neutral Protease: 0.1%； Deoxyribonuclease I: 0.005%；	RPMI	Cerny, D., Haniffa, M., Shin, A., Bigliardi, P., Tan, B., Lee, B., Poidinger, M., Tan, E., Ginhoux, F. and Fink, K.: Selective Susceptibility of Human Skin Antigen Presenting Cells to Productive Dengue Virus Infection., PLoS Pathog 10, e1004548, 2014
人	Human, adult	表皮干细胞 / Epidermal stem cells	Neutral Protease: 2 u/ml； Trypsin: 2.5%	DMEM	Nagel, S., Rohr, F., Weber, C., Kier, J., Siemers, F., Kruse, C., Danner, S., Brandenburger, M. and Matthiessen, A.: Multipotent Nestin-Positive Stem Cells Reside in the Stroma of Human Eccrine and Apocrine Sweat Glands and can be Propagated Robustly In Vitro., PLoS ONE 8, e78365, 2013
人	Human	成纤维细胞 / Fibroblasts	Collagenase animal free: 2%	HBSS	Karumbayaram, S., Lee, P., Azghadi, S., Cooper, A., Patterson, M., Kohn, D., Pyle, A., Clark, A., Byrne, J., Zack, J., Plath, K. and Lowry, W.: From Skin Biopsy to Neurons Through a Pluripotent Intermediate Under Good Manufacturing Practice Protocols, Stem Cells Transl Med 1, 36, 2012
人	Human, female, 18-66 yr	成纤维细胞 / Fibroblasts	Trypsin: 0.2%	DMEM	Huschtscha, L., Napier, C., Noble, J., Bower, K., Au, A., Campbell, H., Braithwaite, A. and Reddel, R.: Enhanced Isolation of Fibroblasts From Human Skin Explants., Biotechniques 53, 239, 2012
人	Human	成纤维细胞和角质形成细胞 / Fibroblasts and keratinocytes	Neutral Protease: 0.5%； Collagenase Type 1: 0.2%	DMEM	Carriel, V., Garzon, I., Jimenez, J., Oliveira, A., Arias-Santiago, S., Campos, A., Sanchez-Quevedo, M. and Alaminos, M.: Epithelial and Stromal Developmental Patterns in a Novel Substitute of the Human Skin Generated with Fibrin-Agarose Biomaterials., Cells Tissues Organs 196, 1-12, 2012
人	Human	表皮 / Epidermal	Collagenase: 0.35%； Neutral Protease: 0.1%	DMEM	Zheng, Y., Nace, A., Chen, W., Watkins, K., Sergott, L., Homan, Y., Vandeberg, J., Breen, M. and Stenn, K.: Mature Hair Follicles Generated from Dissociated Cells: A Universal Mechanism of Folliculoneogenesis., Dev Dyn 239, 2619-26, 2010
人	Human	皮肤和瘢痕疙瘩祖细胞 / Skin and keloid progenitor	Collagenase Type 1: 0.4%； Neutral Protease: 0.3%	PBS	Zhang, Q., Yamaza, T., Kelly, AP, Shi, S., Wang, S., Brown, J., Wang, L., French, S., Shi, S. and Le, A.: Tumor-like Stem Cells Derived from Human Keloid are Governed by the Inflammatory Niche Driven by IL-17/IL-6 Axis., PLoS ONE 4, e7798, 2009
人	Human	成纤维细胞 / Fibroblasts	Collagenase Type 1: 100-320 u/ml	DMEM	Tuan, T., Hwu, P., Ho, W., Yiu, P., Chang, R., Wysocki, A. and Benya, P.: Adenoviral Overexpression and Small Interfering RNA Suppression Demonstrate that Plasminogen Activator Inhibitor-1 Produces Elevated Collagen Accumulation in Normal and Keloid Fibroblasts., Am J Pathol 173, 1311-25, 2008
人	Human, 6-12 year	真皮成纤维细胞 / Dermal fibroblasts	Neutral Protease: 0.1%； Collagenase Type 1: 0.1%	DMEM	Chen, F., Zhang, W., Bi, D., Liu, W., Wei, X., Chen, F., Zhu, L., Cui, L. and Cao, Y.: Clonal Analysis of Nestin(-) Vimentin(+) Multipotent Fibroblasts Isolated from Human Dermis., J Cell Sci 120, 2875, 2007
人	Human	毛囊上皮 / Hair follicular epithelial	Neutral Protease: 1.25%； Trypsin: 0.25%	DMEM/F-12	Yu Hong, Fang Dong, Kumar Suresh M, Li Ling, Nguyen Thiennga K, Acs Geza, Herlyn Meenhard, Xu Xiaowei: Isolation of a novel population of multipotent adult stem cells from human hair follicles, Am J Pathol 168, 1879-88, 2006
人	Human	成纤维细胞 / fibroblasts	Collagenase: 0.5%； Trypsin: 0.25%	DMEM/F12	Clark, R., Chong, B., Mirchandani, N., Yamanaka, K., Murphy, G., Dowgiert, R. and Kupper, T.: A Novel Method for the Isolation of Skin Resident T cells from Normal and Diseased Human Skin., J Invest Dermatol 126, 1059, 2006
人	Human, neonatal	角质形成细胞 / Keratinocytes	Neutral Protease: 0.4%； Collagenase: 0.3%	DMEM	Li, A., Pouliot, N., Redvers, R., and Kaur, P.: Extensive tissue-regenerative capacity of neonatal human keratinocytes stem cells and their progeny, J Clin Invest 113, 390-400, 2004
人	Human	肥大细胞 / Mast cells	Neutral Protease: 0.05%； Collagenase Type 4: 1.0%	RPMI	Babina Magda, Guhl Sven, Starke Andrae, Kirchhof Loreen, Zuberbier Torsten, Henz Beate M: Comparative cytokine profile of human skin mast cells from two compartments--strong resemblance with monocytes at baseline but induction of IL-5 by IL-4 priming, J Leukoc Biol 75, 244-52, 2004
人	Human, adult	真皮成纤维细胞 / Dermal fibroblasts	Collagenase: 0.5-1.0%	DMEM	Wang, H., Van Blitterswijk, C., Bertrand-De Haas, M., Schuurman, A. and Lamme, E.: Improved Enzymatic Isolation of Fibroblasts for the Creation of Autologous Skin Substitutes., In Vitro Cell Dev Biol Anim 40, 268, 2004
人	Human	成纤维细胞 / Fibroblasts	Trypsin: 0.25%	DMEM	Tuan Tai-Lan, Wu Huayang, Huang Eunice Y, Chong Sheree S N, Laug Walter, Messadi Diana, Kelly Paul, Le Anh: Increased plasminogen activator inhibitor-1 in keloid fibroblasts may account for their elevated collagen accumulation in fibrin gel cultures, Am J Pathol 162, 1579-89, 2003
人	Human	角质形成细胞，成纤维细胞，内皮细胞 / Keratinocytes, fibroblasts, endothelial	Neutral Protease: see reference； Collagenase: see reference	see reference	Supp Dorothy M, Wilson-Landy Kaila, Boyce Steven T: Human dermal microvascular endothelial cells form vascular analogs in cultured skin substitutes after grafting to athymic mice, FASEB J 16, 797-804, 2002
人	Human, adult	角质形成细胞 / Keratinocytes	Neutral Protease: 0.25%	PBS	Baudoux, B., Castanares-Zapatero, D., Leclercq-Smekens, M., Berna, N., and Poumay, Y.: The Tetraspanin CD9 Associates with the Integrin a6B4 in Cultured Human Epidermal Keratinocytes and is Involved in Cell Motility, Eur J Cell Biol 79, 41, 2000
人	Human	角质形成细胞 / Keratinocytes	Neutral Protease: 0.25%； Trypsin: 0.17%； Thermolysin: 0.05%；	see reference	Hybbinette S, Bostrom M, Lindberg K: Enzymatic dissociation of keratinocytes from human skin biopsies for in vitro cell propagation, Exp Dermatol 8, 30-8, 1999
人	Human	人皮肤肥大细胞 / Human skin mast cells	Neutral Protease: 0.1%； Collagenase Type 1: see reference	PBS	Grutzkau A, Kruger-Krasagakes S, Baumeister H, Schwarz C, Kogel H, Welker P, Lippert U, Henz BM, Moller A: Synthesis, storage, and release of vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) by human mast cells: implications for the biological significance of VEGF206, Mol Biol Cell 9, 875-84, 1998