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：
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 |
| 人 | Human | 表皮 / Epidermal | Collagenase Type 2: 0.1%； Neutral Protease: 0.5-1.0% |
PBS | Reece J C, Handley A J, Anstee E J, Morrison W A, Crowe S M, Cameron P U: HIV-1 selection by epidermal dendritic cells during transmission across human skin, J Exp Med 187, 1623-31, 1998 |
| 人 | Human | 角质形成细胞 / Keratinocytes | Trypsin: 0.05% | Dulbecco's PBS | Judd, D., Battista, P., and Behm, D.: Culture of Human Keratinocytes in Defined Serum-Free Medium, Focus 19 (1), 2, 1997 |
| 人 | Human | 表皮加真皮 / Epidermis plus dermis | Trypsin: 0.125% | DMEM, M199 | Harley, C., and Sherwood, S.: Methods in Molecular Biology, Basic Cell Culture Protocols, 2nd ed. Vol. 75,Pollard, J., and Walker, J., Humana Press, 23, 1997 |
| 人 | Human | 角质形成细胞 / Keratinocytes | Trypsin: 0.25% | DMEM | Regnier, M: Culture of Human Karatinocytes,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons Ltd, 11B:4.1, 1995 |
| 人 | Human, adult, female, breast skin | 角质形成细胞 / Keratinocytes | Trypsin: 0.25% | PBS | Hirel, B., Chesne, C., Pailgheret, J., and Guillouzo, A.: Expression of Differentiation Markers in Human Adult Keratinocytes Cultured in Submerged Conditions, In Vitro Cell Dev Biol 30A, 372, 1994 |
| 人 | Human, neonatal foreskin | 成纤维细胞 / Fibroblasts | Trypsin: 0.25% | DMEM | Hansbrough, J., Cooper, M., Cohen, R., Spielvogel, R., Greenleaf, G., Bartel, R., and Naughton, G.: Evaluation of a Biodegradable Matrix Containing Cultured Human Fibroblasts as a Dermal Replacement Beneath Meshed Skin Grafts on Athymic Mice, Surgery 111 (1), 438, 1992 |
| 人 | Human, 18-50 years | 汗管 / Sweat duct | Collagenase: 0.03% | MEM | Bell, C. and Quinton, P.: Effects of Media Buffer Systems on Growth and Electrophysiologic Characteristics of Cultured Sweat Duct Cells, In Vitro Cell Dev Biol 27, 47, 1991 |
| 人 | Human | 成纤维细胞 / Fibroblasts | Trypsin: 0.25% | CMF solution | Limat, A., Hunziker, T., Boillat, C., Noser, F., and Wiesmann, U.: Postmitotic Human Dermal Fibroblasts Preserve Intact Feeder Properties for Epithelial Cell Growth After Long-Term Cryopreservation, In Vitro Cell Dev Biol 26, 709, 1990 |
| 人 | Human | 汗腺 / Sweat gland | Collagenase Type 2: 0.015% | (see reference) | Krouse, M., Hagiqara, G., Chen, J., Lewiston, N., and Wine, J.: Ion Channels in Normal Human and Cystic Fibrosis Sweat Gland Cells, Am J Physiol 257, C129, 1989 |
| 人 | Human, ages 16-30 | 人体汗管 / Human sweat duct | Collagenase Type 2: 0.2% | RPMI 1640 | Pedersen, P.: Human Sweat Duct Cells in Primary Culture. Basic Bioelectric Properties of Cultures Derived From Normals and Patients with Cystic Fibrosis, In Vitro Cell Dev Biol 25 (4), 342, 1989 |
| 人 | Human | 黑色素细胞，皮肤/包皮 / Melanocytes, skin/ foreskin | Trypsin: 0.25% | PBS | Peacocke, M., Yaar, M., Mansur, C., Chao, M., and Gilchrest, B.: Induction of Nerve Growth Factor Receptors on Cultured Human Melanocytes, Proc Natl Acad Sci U S A 85, 5282, 1988 |
| 人 | Human | 人体汗腺 / Human sweat glands | Collagenase: 0.2% | HBSS | Lee, C., Carpenter, F., Coaker, T., and Kealey, T.: The Primary Culture of Epithelia From the Secretory Coil and Collecting Duct of Normal Human and Cystic Fibrotic Eccrine Sweat Glands, J Cell Sci 83, 103, 1986 |
| 人 | Human | 角质形成细胞 / Keratinocytes | Trypsin: 0.25% | DMEM | Dover, R. and Potten, C.: Cell Cycle Kinetics of Cultured Human Epidermal Keratinocytes, J Invest Dermatol 80, 423, 1983 |
| 人 | Human, adult | 表皮角质形成细胞 / Epidermal keratinocytes | Trypsin: 0.25%；
Collagenase: 0.2%； Deoxyribonuclease I: 0.001%； |
Eagle's MEM | Alitalo K, Kuismanen E, Myllyla R, Kiistala U, Asko-Seljavaara S, Vaheri A: Extracellular matrix proteins of human epidermal keratinocytes and feeder 3T3 cells, J Cell Biol 94, 497-505, 1982 |
| 人 | Human, fetal | 平滑肌，成纤维细胞 / Smooth muscle, fibroblasts | Trypsin: 0.055% | DMEM | Davies, P. and Kerr, C.: Modification of LDL Metabolism by Growth Factors in Cultured Vascular Cells and Human Skin Fibroblasts, Biochim Biophys Acta 712, 26, 1982 |
| 人 | Human, 18-30 years, male, female | 成纤维细胞 / Fibroblasts | Trypsin: 0.1% | HBSS | McCoy, B., Galdun, J., and Cohen, I.: Effects of Density and Cellular Aging On Collagen Synthesis and Growth Kinetics in Keloid and Normal Skin Fibroblasts, In Vitro 18 (1), 79, 1982 |
| 人 | Human, newborn | 角质形成细胞 / Keratinocytes | Trypsin: 0.3% | DMEM | Liu, S., Eaton, M., and Karasek, M.: Growth Characteristics of Human Epidermal Keratinocytes from Newborn Foreskin in Primary and Serial Cultures, In Vitro 15 (10), 813, 1979 |
| 人 | Human | 角质形成细胞 / Keratinocytes | Trypsin: 0.25% | (see reference) | Rheinwald, J., and Green, H.: Serial Cultivation of Strains of Human Epidermal Kerotinocytes: The Formation of Keratinizing Colonies from Single Cells, Cell 6, 331, 1975 |
| 小鼠 | Mouse | 角质形成细胞 / Keratinocytes | Collagenase Type 4: 0.2%；
Deoxyribonuclease I: 0.005%； Neutral Protease: 5 u/ml； |
DMEM | Spidale, N., Malhotra, N., Frascoli, M., Sylvia, K., Miu, B., Freeman, C., Stadinski, B., Huseby, E. and Kang, J.: Neonatal-Derived IL-17 Producing Dermal gd T Cells are Required to Prevent Spontaneous Atopic Dermatitis, Elife 9, 5118, 2020 |
| 小鼠 | Mouse | 角质形成细胞 / Keratinocytes | Trypsin: 0.25%；
Collagenase Type 1: 0.12%； Collagenase Type 2: 0.05%； Collagenase Type 4: 0.05%； Hyaluronidase: 0.1%； |
Low calcium containing media | Blanco, S., Bandiera, R., Popis, M., Hussain, S., Lombard, P., Aleksic, J., Sajini, A., Tanna, H., Cortes-Garrido, R., Gkatza, N., Dietmann, S. and Frye, M.: Stem Cell Function and Stress Response are Controlled by Protein Synthesis., Nature 534, 335-40, 2016 |
| 小鼠 | Mouse | 真皮成纤维细胞 / Dermal fibroblasts | Elastase: 0.012%； Collagenase Type 4: 0.4% |
DMEM | Rinkevich, Y., Walmsley, G., Hu, M., Maan, Z., Newman, A., Drukker, M., Januszyk, M., Krampitz, G., Gurtner, G., Lorenz, H., Weissman, I. and Longaker, M.: Identification and Isolation of a Dermal Lineage with Intrinsic Fibrogenic Potential., Science 348, aaa2151, 2015 |
| 小鼠 | Mouse | 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 |
| 小鼠 | Mouse | 表皮 / Epidermal | Neutral Protease: 0.5%； Collagenase Type 4: 0.1% |
DMEM | Gu, D., Fan, Q and Xie, J.: Cell Population Analyses During Skin Carcinogenesis., J Vis Exp , e50311, 2013 |
| 小鼠 | Mouse | 上皮干细胞 / Epithelial stem | Trypsin: 0.25%； Collagenase Type 1: 0.4% |
HBSS | Jensen, U., Ghazizadeh, S. and Owens, D.: Isolation and Characterization of Cutaneous Epithelial Stem Cells., Methods Mol Biol 989, 61-9, 2013 |
| 小鼠 | Mouse | 桅杆 / Mast | Collagenase Type 4: 0.2% | IMDM | Antsiferova, M., Martin, C., Huber, M., Feyerabend, T., Forster, A., Hartmann, K., Rodewald, H., Hohl, D. and Werner, S.: Mast Cells are Dispensable for Normal and Activin-Promoted Wound Healing and Skin Carcinogenesis., J Immunol 191, 6147-55, 2013 |
| 小鼠 | Mouse | 真皮成纤维细胞 / Dermal fibroblasts | Collagenase: 400 u/ml | AminoMAX II | Scheibye-Knudsen, M., Ramamoorthy, M., Sykora, P., Maynard, S., Lin, P., Minor, R., Wilson, D., Cooper, M., Spencer, R., de Cabo, R., Croteau, D. and Bohr, V.: Cockayne Syndrome Group B Protein Prevents the Accumulation of Damaged Mitochondria by Promoting Mitochondrial Autophagy., J Exp Med 209, 855, 2012 |
| 小鼠 | Mouse | 表皮和真皮 / Epidermal and dermal | Collagenase Type 4: 0.18%； Collagenase/Dispase: 0.18% |
PBS | King, I., Kroenke, M. and Segal, B.: GM-CSF-Dependent, CD103+ Dermal Dendritic Cells Play a Critical Role in Th Effector Cell Differentiation After Subcutaneous Immunization., J Exp Med 207, 953, 2010 |
| 小鼠 | Mouse, 2-3 day | 真皮成纤维细胞 / Dermal fibroblasts | Collagenase Type 2: 0.04%； Trypsin: 0.025% |
DME | Farina, G., York, M., Di Marzio, M., Collins, C., Meller, S., Homey, B., Rifkin, I., Marshak-Rothstein, A., Radstake, T. and Lafyatis, R.: Poly(I:C) Drives Type I IFN- and TGFβ-Mediated Inflammation and Dermal Fibrosis Simulating Altered Gene Expression in Systemic Sclerosis., J Invest Dermatol 130, 2583-93, 2010 |
| 小鼠 | Mouse | 真皮 / Dermal | Collagenase Type 3: 0.3%； Deoxyribonuclease I: 0.0005% |
HBSS | Eidsmo, L., Allan, R., Caminschi, I., Van Rooijen, N., Heath, W. and Carbone, F.: Differential Migration of Epidermal and Dermal Dendritic Cells During Skin Infection., J Immunol 182, 3165, 2009 |
| 小鼠 | Mouse, neonatal | 真皮 / Dermal | Collagenase: 0.35%； Deoxyribonuclease I: see reference |
PBS | Crigler Lauren, Kazhanie Amita, Yoon Tae-Jin, Zakhari Julia, Anders Joanna, Taylor Barbara, Virador VictoriaM: Isolation of a mesenchymal cell population from murine dermis that contains progenitors of multiple cell lineages, FASEB J 21, 2050-63, 2007 |
| 小鼠 | mouse | 真皮成纤维细胞 / Dermal fibroblasts | Trypsin: 0.25%； Collagenase Type 1: 0.25% |
DMEM | Baxter Ruth M, Crowell Thomas P, McCrann Margaret E, Frew Erica M, Gardner Humphrey: Analysis of the tight skin (Tsk1/+) mouse as a model for testing antifibrotic agents, Lab Invest 85, 1199-209, 2005 |
| 小鼠 | Mouse, neonatal | 微血管内皮 / Microvascular endothelial | Neutral Protease: 0.005%； Collagenase Type 1: 4% |
DMEM | Cha, S., Talavera, D., Demir, E., Nath, A. and Sierra-Honigmann, M.: A Method of Isolation and Culture of Microvascular Endothelial Cells from Mouse Skin., Microvasc Res 70, 198, 2005 |
| 小鼠 | Mouse, male 6-24 week | 皮侧人群 / Skin side population | Collagenase Type 4: 0.2%； Neutral Protease: 1.2 u/ml |
PBS | Montanaro F, Liadaki K, Volinski J, Flint A, and Kunkel LM.: Skeletal muscle engraftment potential of adult mouse skin side population cells, Proc Natl Acad Sci U S A 100, 9336, 2003 |
| 小鼠 | Mouse | 耳表皮 / Ear epidermal | Trypsin: 0.1%； Collagenase: 0.2% |
PBS | Takanami-Ohnishi Yoko, Amano Shinya, Kimura Sadao, Asada Sachie, Utani Atsushi, Maruyama Masumi, Osada Hiroyuki, Tsunoda Hajime, Irukayama-Tomobe Yoko, Goto Katsutoshi, Karin Michael, Sudo Tatsuhiko, Kasuya Yoshitoshi: Essential role of p38 mitogen-activated protein kinase in contact hypersensitivity, J Biol Chem 277, 37896-903, 2002 |
| 小鼠 | Mouse, 3-7 month | 成纤维细胞，系膜，平滑肌 / Fibroblasts, mesangial, smooth muscle | Trypsin: 0.25%；
Collagenase: see reference； Soybean Trypsin Inhibitor: 0.05%； |
DMEM | Bradshaw AD, Francki A, Motamed K, Howe C, Sage EH: Primary mesenchymal cells isolated from SPARC-null mice exhibit altered morphology and rates of proliferation, Mol Biol Cell 10, 1569-79, 1999 |
| 猪 | Porcine, 3-4 month | 滑膜和皮肤干细胞 / Synovial membrane and skin stem | Collagenase: 0.2% | DMEM | Ando, W., Kutcher, J., Krawetz, R., Sen, A., Nakamura, N., Frank, C. and Hart, D.: Clonal Analysis of Synovial Fluid Stem Cells to Characterize and Identify Stable Mesenchymal Stromal Cell/Mesenchymal Progenitor Cell Phenotypes in a Porcine Model: A Cell Source with Enhanced Commitment to the Chondrogenic Lineage., Cytotherapy 16, 776, 2014 |
| 猪 | Porcine, 2-6 month | 角质形成细胞 / Keratinocytes | Neutral Protease: 0.25% | Dulbecco-Vogt MEM | Regauer S, Compton C: Cultured porcine epithelial grafts: an improved method, J Invest Dermatol 94, 230-4, 1990 |
| 大鼠 | Rat, Wistar, 4-8 day | 真皮成纤维细胞和角质形成细胞 / Dermal fibroblasts and keratinocytes | Trypsin: 0.25% | Ham's F-12 | Sugihara, H and Toda, S: Primary Tissue Intact and Dissociated Cell Culture,Cell & Tissue Culture: Laboratory Procedures Vol. 1,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 3A:2.1, 1995 |
| 大鼠 | Rat, SD, male, 40-60 days | 皮脂腺 / Sebaceous | Trypsin: 0.2% | DMEM with FBS | Laurent, S., Mednieks, M., and Rosenfield, R.: Growth of Sebaceous Cells in Monolayer Culture, In Vitro Cell Dev Biol 28, 83, 1992 |
| 大鼠 | Rat, neonatal, 12-24 hr post partum | 成纤维细胞 / Fibroblasts | Trypsin: 0.2% | HEPES buffered DMEM | Acheson, A., Barker, P., Alderson, R., Miller, F., and Murphy, R.: Detection of Brain-Derived Neurotrophic Factor-like Activity in Fibroblasts and Schwann Cells: Inhibition by Antibodies to NGF, Neuron 7, 265, 1991 |
| 大鼠 | Rat, albino, one day old, CFN | 角质形成细胞 / Keratinocytes | Trypsin: 1% | EBSS | Vaughan, F., Gray, R., and Bernstein, I.: Growth and Differentiation of Primary Rat Keratinocytes on Synthetic Membranes, In Vitro Cell Dev Biol 22 (3), 141, 1986 |
