世界*实验材料供应商 Worthington Biochemical 正式授权上海起发为其中国代理， Worthington Biochemical 在一直是行业的*,一直为广大科研客户提供zui为的产品和服务,上海起发一直秉承为中国科研客户带来的产品，的服务，签约 Worthington Biochemical 就是为了给广大科研客户带来更加完善的产品和服务,您的满意将是我们zui大的收获
Worthington Biochemical 中国代理, Worthington Biochemical 上海代理， Worthington Biochemical 北京代理，Worthington Biochemical 广东代理， Worthington Biochemical 江苏代理Worthington Biochemical 湖北代理,Worthington Biochemical 天津,Worthington Biochemical 黑龙江代理,Worthington Biochemical 内蒙古代理,Worthington Biochemical 吉林代理,Worthington Biochemical 福建代理, Worthington Biochemical 江苏代理, Worthington Biochemical 浙江代理,Worthington Biochemical 四川代理,
Worthington组织消化指南
引言
组织消化酶主要用于组织培养和细胞生物学研究的原代细胞的分离和收集。尽管这些消化酶已经普遍应用多年，但是对其组织消化和细胞收集的作用机制却没有全面的理解。 由此导致的结果便是选择一种消化方法大多是随机选择或基于过去的经验，而不是在*理解消化原理的基础上选择的消化方案，以及如何根据具体实验目的修正消化方案以获得更好的实验结果。
细胞分离程序的目的是zui大程度的获得功能活性的单个细胞，而影响结果的原因有很多，包括（不限于）以下：
1、组织类型
2、物种来源
3、动物年龄
4、遗传改造情况（如knockouts）
5、所用的消化培养液
6、所用的酶
7、消化酶中的杂质
8、所用消化酶的工作浓度
9、温度
10、孵育时间
通常前四个原因没办法选择，而剩下的原因一般由经验决定以获取zui适实验结果。
通过查阅文献来选择*组织消化酶和zui适消化条件的研究人员通常被互相矛盾的实验数据搞得一头雾水。这些巨大差异来源于组织胞外基质的复杂而动态的特性，以及过去常用的粗制、未提纯的组织分离酶。另外，组织胞外基质由各种蛋白、糖蛋白、脂类和糖脂组成，这些成分的组成比例在不同物种、不同组织、甚至不同发育（年龄）之间存在明显的差异，这些因素都极大的影响了消化方案的制定。目前通常使用的粗制酶，如链霉蛋白酶、NF1:250、以及含有几种其他不同浓度蛋白酶的胶原酶、以及各种多糖酶、核酶和脂酶。
本指南总结了目前常用组织消化酶的zui适消化和细胞收集知识和方法，描述了标准的实验步骤， 以便于制定一个*的细胞分离方案，同时列举了一些相关的参考文献。
下表列出多种组织的适宜消化酶（含参考文献，以组织类型和物种分类，详见链接）
脂肪组织 | 肾上腺组织 | 骨组织 | 脑组织 |
软骨组织 | 结肠组织 | 内皮组织 | 上皮组织 |
眼 | 心脏 | 肠 | 肾 |
肝 | 肺 | 淋巴结 | 乳腺 |
其它组织 | 其他组织 | 肌组织 | 神经组织 |
胰腺 | 腮腺 | 垂体 | 前列腺 |
生殖 | 动物鳞片 | 皮肤 | 脾脏 |
干细胞 | 胸腺 | 甲状腺/甲状旁腺 | 扁桃体 |
肿瘤 |
|
|
|
注：使用Worthington公司消化酶的文献不限于上表链接中所列，一般而言，Worthington公司的组织消化酶可用于大多组织类型的消化。
皮肤组织消化手册
| 物种 | 详细来源 | 细胞名称 | 消化用酶 | 培养基 | 参考文献 |
| 犬 | 犬 | Epidermal | Collagenase: 0.35% | 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 |
| Neutral Protease: 0.1% | |||||
| 青蛙 | 青蛙 | 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 | 山羊 | Dermis isolated aggrecan sensitive | Neutral Protease: 0.5% | 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 |
| Collagenase Type 2: 200 u/ml | |||||
| 人类 | 人类 | Skin | Neutral Protease: 5 u/ml | 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 |
| Collagenase Type 3: 0.3% | |||||
| Deoxyribonuclease I: 0.005% | |||||
| 人类 | 人类 | 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 Reports3, 250, 2014 |
| 人类 | 人类, 成年的 | Epidermal stem cells | Neutral Protease: 2 u/ml | 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 |
| Trypsin: 2.5% | |||||
| 人类 | 人类 | 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 |
| 人类 | 人类, 女性, 18-66 岁 | 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 |
| 人类 | 人类 | Epidermal | Collagenase: 0.35% | 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 |
| Neutral Protease: 0.1% | |||||
| 人类 | 人类 | skin and keloid progenitor cells | Collagenase Type 1: 0.4% | 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 |
| Neutral Protease: 0.3% | |||||
| 人类 | 人类 | 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 |
| 人类 | 人类, 6-12 岁 | Dermal fibroblasts | Neutral Protease: 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 Sci120, 2875, 2007 |
| Collagenase Type 1: 0.1% | |||||
| 人类 | 人类 | Hair follicular epithelial | Neutral Protease: 1.25% | 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 |
| Trypsin: 0.25% | |||||
| 人类 | 人类 | fibroblasts | Collagenase: 0.5% | 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 |
| Trypsin: 0.25% | |||||
| 人类 | 人类, 新生的 | Keratinocytes | Neutral Protease: 0.4% | 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 |
| Collagenase: 0.3% | |||||
| 人类 | 人类 | Mast cells | Neutral Protease: 0.05% | 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 |
| Collagenase Type 4: 1.0% | |||||
| 人类 | 人类, 成年的 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类 | Keratinocytes, fibroblasts, endothelial | Neutral Protease: 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 |
| Collagenase: see reference | |||||
| 人类 | 人类, 成年的 | 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 |
| 人类 | 人类 | Keratinocytes | Neutral Protease: 0.25% | 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 |
| Trypsin: 0.17% | |||||
| Thermolysin: 0.05% | |||||
| 人类 | 人类 | Human skin mast cells | Neutral Protease: 0.1% | 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 |
| Collagenase Type 1: see reference | |||||
| 人类 | 人类 | Epidermal | Collagenase Type 2: 0.1% | 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 |
| Neutral Protease: 0.5-1.0% | |||||
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| Abdomen or inner forearm | |||||
| 人类 | 人类 | 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 |
| 人类 | 人类, 成年的, 女性, 乳房皮肤 | 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 |
| 人类 | 人类, 新生儿包皮 | Fibroblasts | Trypsin: 0.25% | DMEM | Hansbrough, J., Cooper, M., Cohen, R., Spielvogel, R., Greenleaf, G., Bar, 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 |
| 人类 | 人类, 18-50 岁 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类, 年龄16-30 | 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 |
| (see reference) | |||||
| 人类 | 人类 | 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 |
| 人类 | 人类 | 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 |
| 人类 | 人类 | Keratinocytes | Trypsin: 0.25% | DMEM | Dover, R. and Potten, C.: Cell Cycle Kinetics of Cultured Human Epidermal Keratinocytes, J Invest Dermatol 80, 423, 1983 |
| 人类 | 人类, 成年的 | Epidermal keratinocytes | Trypsin: 0.25% | 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 |
| Collagenase: 0.2% | |||||
| Deoxyribonuclease I: 0.001% | |||||
| 人类 | 人类, fetal | 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 |
| 人类 | 人类, 18-30岁, 男性, 女性 | 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 |
| 人类 | 人类, 新生儿 | 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 |
| 人类 | 人类 | Keratinocytes | Trypsin: 0.25% | (see reference) | Rheinwald, J., and Green, H.: Serial C*tion of Strains of Human Epidermal Kerotinocytes: The Formation of Keratinizing Colonies from Single Cells, Cell 6, 331, 1975 |
| 小鼠 | 小鼠 | Epidermal | Neutral Protease: 0.5% | DMEM | Gu, D., Fan, Q and Xie, J.: Cell Population Analyses During Skin Carcinogenesis., J Vis Exp , e50311, 2013 |
| Collagenase Type 4: 0.1% | |||||
| 小鼠 | 小鼠 | Epithelial Stem | Trypsin: 0.25% | HBSS | Jensen, U., Ghazizadeh, S. and Owens, D.: Isolation and Characterization of Cutaneous Epithelial Stem Cells., Methods Mol Biol 989, 61-9, 2013 |
| Collagenase Type 1: 0.4% | |||||
| 小鼠 | 小鼠 | 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 | 小鼠 | Epidermal and dermal | Collagenase Type 4: 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 |
| Collagenase/Dispase: 0.18% | |||||
| 小鼠 | 小鼠, 2-3 天 | Dermal fibroblasts | Collagenase Type 2: 0.04% | 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 TGF尾-Mediated Inflammation and Dermal Fibrosis Simulating Altered Gene Expression in Systemic Sclerosis., J Invest Dermatol 130, 2583-93, 2010 |
| Trypsin: 0.025% | |||||
| 小鼠 | 小鼠 | Dermal | Collagenase Type 3: 0.3% | 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 |
| Deoxyribonuclease I: 0.0005% | |||||
| 小鼠 | 小鼠, 新生的 | Dermal | Collagenase: 0.35% | 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 |
| Deoxyribonuclease I: see reference | |||||
| 小鼠 | 小鼠 | Dermal fibroblasts | Trypsin: 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 |
| Collagenase Type 1: 0.25% | |||||
| 小鼠 | 小鼠, 新生的 | Microvascular endothelial | Neutral Protease: 0.005% | 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 |
| Collagenase Type 1: 4% | |||||
| 小鼠 | 小鼠, male 6-24 week | Skin side population | Collagenase Type 4: 0.2% | 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 |
| Neutral Protease: 1.2 u/ml | |||||
| 小鼠 | 小鼠 | Ear epidermal | Trypsin: 0.1% | 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 |
| Collagenase: 0.2% | |||||
| 小鼠 | 小鼠, 3-7 月 | Fibroblasts, mesangial, smooth muscle | Trypsin: 0.25% | 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 |
| Collagenase: see reference | |||||
| Soybean Trypsin Inhibitor: .05% | |||||
| 猪 | 猪, 3-4 月 | Synovial membrane and skin stem cells | 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 | 猪, 2-6 月 | 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 | Wistar大鼠, 4-8 天 | 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 | SD大鼠, 雄性, 40-60 天 | 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 |
| PBS | |||||
| 大鼠 | 大鼠, 新生的, 产后的12-24 小时 | 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 | 大鼠, 白化变种, 一天大, CFN | 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 |
我们公司zui大优势是强大的采购，
1：基本什么都能进口，血清，抗体，耗材，还有部分限制进口的，
2：货品全，现经营过700多个品牌，基本所有生物试剂耗材都可以进口，特别是冷偏的产品那就更有优势，
3：提供加急服务，一般1-2周到货，超过时限加急费全免
4：价格公道，绝大部分价格有优势，当然不能保证100%产品都是,因为意味着没有服务.
5：良好的信誉，大部分客户我们提供货到付款服务，客户包括清华，北大 交大 复旦，中山等100多所大学，ROCHE，阿斯利康，国药 ，fisher等500多家公司
6：我们还是Santa,Advanced Biotechnologies Inc, Worthington Biochemical ,bangs,BBInternational,crystalchem,dianova,FD Neurotechnologies,Inc. Dako,Inc, GePromegaridege, Glycotope Biotechnology GmbH; iduron,Innovative Research of America, Ludger, neuroprobe,Worthington Biochemical , Polysciences,prospecbi, QA-BIO,quickzyme,FULLERLABS,INC,sterlitech；sysy,TriLink BioTechnologies,Inc；worthington-biochem,zyagen等几十家国外公司授权代理。
7：我们还是invitrogen,qiagen,Worthington Biochemical am,sigma;Promega,roche,merck, rnd,BD, GE,pierce,BioLegend等*批发，欢迎合作。
