worthington 皮肤组织分离指南

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:

worthington 胰腺组织分离指南

 

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

 

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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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