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2016/2/16 16:02:27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
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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。
干细胞组织消化手册
物种 | 详细来源 | 细胞名称 | 消化用酶 | 培养基 | 参考文献 |
鸟 | 人 | Adipose derived stem cells | Collagenase Type 1: 0.1% | DMEM | Tandon, N., Goh, B., Marsano, A., Chao, P., Montouri-Sorrentino, C., Gimble, J. and Vunjak-Novakovic, G.: Alignment and Elongation of Human Adipose-Derived Stem Cells in Response to Direct-Current Electrical Stimulation., Conf Proc IEEE Eng Med Biol Soc 2009, 6517-21, 2009 |
犬 | 犬 | Amniotic membrane MSC | Trypsin: 0.25% | LG-DMEM | Park, S., Seo, M., Kim, H. and Kang, K.: Isolation and Characterization of Canine Amniotic Membrane-Derived Multipotent Stem Cells., PLoS ONE 7, e44693, 2012 |
Collagenase Type 1: 0.2% | |||||
犬 | 犬, 20-25 公斤 | Adipose stem cells | Collagenase: see reference | Media-199 | Fischer, L., McIlhenny, S., Tulenko, T., Golesorkhi, N., Zhang, P., Larson, R., Lombardi, J., Shapiro, I. and DiMuzio, P.: Endothelial Differentiation of Adipose-Derived Stem Cells: Effects of Endothelial Cell Growth Supplement and Shear Force., J Surg Res 152, 157, 2009 |
马 | 马 | Adipose derived stem cells | Collagenase Type 1: 0.1% | PBS | Vidal, M., Robinson, S., Lopez, M., Paulsen, D., Borkhsenious, O., Johnson, J., Moore, R. and Gimble, J.: Comparison of Chondrogenic Potential in Equine Mesenchymal Stromal Cells Derived from Adipose Tissue and Bone Marrow., Vet Surg 37, 713, 2008 |
马 | 马, 1-5 岁 | Adipose derived stem cells | Collagenase Type 1: 0.1% | PBS | Vidal, M., Kilroy, G., Lopez, M., Johnson, J., Moore, R. and Gimble, J.: Characterization of Equine Adipose Tissue-Derived Stromal Cells: Adipogenic and Osteogenic Capacity and Comparison with Bone Marrow-Derived Mesenchymal Stromal Cells., Vet Surg 36, 613, 2007 |
人类 | 人类 | Mesenchymal Stromal Cells | Collagenase Type 4: 0.2% | PBS | Choudhery,M, Badowski, M, Muise, A and Harris, D: Effect of Mild Heat Stress on the Proliferative and Differentiative Ability of Human Mesenchymal Stromal Cells., Cytotherapy17, 359-68, 2015 |
人类 | 人类 | Adipose derived stem cells | Collagenase Type 2: 0.1% | HBSS | Satish, L., Krill-Burger, J., Gallo, P., Etages, S., Liu, F., Philips, B., Ravuri, S., Marra, K., LaFramboise, W., Kathju, S. and Rubin, J.: Expression Analysis of Human Adipose-Derived Stem Cells During In Vitro Differentiation to an Adipocyte Lineage., BMC Med Genomics 8, 41, 2015 |
人类 | 人类 | Dental mesenchymal stem | Collagenase Type 1: 0.3% | DMEM | Wu, Z., Wang, J., Dong, R., Wang, L., Fan, Z., Liu, D. and Wang, S.: Depletion of MEIS2 Inhibits Osteogenic Differentiation Potential of Human Dental Stem Cells., Int J Clin Exp Med8, 7220-30, 2015 |
Neutral Protease: 0.4% | |||||
人类 | 人类 | Decidua Mesenchymal Stem | Trypsin: 0.25% | PBS | Kusuma, G., Menicanin, D., Gronthos, S., Manuelpillai, U., Abumaree, M., Pertile, M., Brennecke, S. and Kalionis, B.: Ectopic Bone Formation by Mesenchymal Stem Cells Derived from Human Term Placenta and the Decidua., PLoS ONE 10, e0141246, 2015 |
Collagenase Type 1: 1.0% | |||||
Deoxyribonuclease I: 0.005% | |||||
人类 | 人类, 成年人 | Adipose derived mesenchymal stem | Collagenase Type 2: 0.1% | see reference | Al-Saqi, S., Saliem, M., Asikainen, S., Quezada, H., Ekblad, A., Hovatta, O., Le Blanc, K., Jonasson, A. and Gotherstrom, C.: Defined Serum-Free Media for In Vitro Expansion of Adipose-Derived Mesenchymal Stem Cells., Cytotherapy 16, 915, 2014 |
人类 | 人类, 成年人 | Adipose derived stem cells | Collagenase Type 1: 0.15% | DMEM | Koellensperger, E., Bollinger, N., Dexheimer, V., Gramley, F., Germann, G. and Leimer, U.: Choosing the Right Type of Serum for Different Applications of Human Adipose Tissue-Derived Stem Cells: Influence on Proliferation and Differentiation Abilities., Cytotherapy 16, 789, 2014 |
人类 | 人类 | Umbilical mesenchymal stem | Collagenase Type 2: 0.1% | DMEM | Hang, H., Yu, Y., Wu, N., Huang, Q., Xia, Q. and Bian, J.: Induction of Highly Functional Hepatocytes from Human Umbilical Cord Mesenchymal Stem Cells by HNF4伪 Transduction.,PLoS ONE 9, e104133, 2014 |
Trypsin: 0.25% | |||||
人类 | 人类 | Cardiac stem | Collagenase Type 2: 0.25% | Joklik modified Eagle抯 | Avolio, E., Gianfranceschi, G., Cesselli, D., Caragnano, A., Athanasakis, E., Katare, R., Meloni, M., Palma, A., Barchiesi, A., Vascotto, C., Toffoletto, B., Mazzega, E., Finato, N., Aresu, G. and Livi, U.: Ex vivo Molecular Rejuvenation Improves the Therapeutic Activity of Senescent Human Cardiac Stem Cells in a Mouse Model of Myocardial Infarction., Stem Cells 32, 2373, 2014 |
人类 | 人类 | Adipose stromal stem | Collagenase animal free: 200 u/ml | DMEM/Hams F-12 | Carvalho, P., Gimble, J., Dias, I., Gomes, M. and Reis, R.: Xenofree Enzymatic Products for the Isolation of Human Adipose-Derived Stromal/Stem Cells., Tiss Eng 19, 473-8, 2013 |
人类 | 人类, 成年人 | 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 | Dental pulp stem | Collagenase Type 1: 0.3% | DMEM/F12 | Bonnamain, V., Thinard, R., Sergent-Tanguy, S., Huet, P., Bienvenu, G., Naveilhan, P., Farges, J. and Alliot-Licht, B.: Human Dental Pulp Stem Cells Cultured in Serum-Free Supplemented Medium., Front Physiol 4, 357, 2013 |
Neutral Protease: 0.4% | |||||
人类 | 人类 | Umbilical Cord Mesenchymal Stem | Collagenase: 0.1-0.2% | DMEM | Han, Y., Tao, R., Sun, T., Chai, J., Xu, G. and Liu, J.: Optimization of Human Umbilical Cord Mesenchymal Stem Cell Isolation and Culture Methods., Cytotechnology 65, 819, 2013 |
Trypsin: 0.1% | |||||
人类 | 人类 | Dental pulp derived stem cell | Collagenase Type 1: 0.3% | DMEM | Sakai, K., Yamamoto, A., Matsubara, K., Nakamura, S., Naruse, M., Yamagata, M., Sakamoto, K., Tauchi, R., Wakao, N., Imagama, S., Hibi, H., Kadomatsu, K., Ishiguro, N. and Ueda, M: Human Dental Pulp-Derived Stem Cells Promote Locomotor Recovery After Complete Transection of the Rat Spinal Cord by Multiple Neuro-Regenerative Mechanisms., J Clin Invest 122, 80, 2012 |
Neutral Protease: 0.4% | |||||
人类 | 人类 | Adipose-derived stem cells | Collagenase Type 1: 0.1% | PBS | Cervelli, V., Scioli, M., Gentile, P., Doldo, E., Bonanno, E., Spagnoli, L. and Orlandi, A.:Plaet-Rich Plasma Greatly Potentiates Insulin-Induced Adipogenic Differentiation of Human Adipose-Derived Stem Cells Through a Serine/Threonine Kinase Akt-dependent Mechanism and Promotes Clinical Fat Graft Maintenance., Stem Cells Transl Med 1, 206-20, 2012 |
人类 | 人类 | Mesenchymal stem | Collagenase Type 1: 0.4% | DMEM/F12 | Shalini, V., Pratheep, S., Muhammad, A., Sharmili, V., Elizabeth, G. and Rajesh, R.: Generation and Characterisation of Human Mesenchymal Stem Cells Derived From Umbilical Cord and Placenta, Regenerative Research 1, 48, 2012 |
Deoxyribonuclease I: 0.01% | |||||
人类 | 人类 | Wharton jelly mesenchymal stem | Collagenase Type 2: 0.1% | DMEM | Salehinejad, P., Alitheen, N., Ali, A., Omar, A., Mohit, M., Janzamin, E., Samani, F., Torshizi, Z. and Nematollahi-Mahani, S.: Comparison of Different Methods for the Isolation of Mesenchymal Stem Cells from Human Umbilical Cord Wharton's Jelly., In Vitro Cell Dev Biol Anim 48, 75, 2012 |
Trypsin: 0.25% | |||||
人类 | 人类 | Placental Mesenchymal Stem | Collagenase Type 2: 0.4% | DMEM | Vellasamy, S., Sandrasaigaran, P., Vidyadaran, S., George, E. and Ramasamy, R.:Isolation and Characterisation of Mesenchymal Stem cells Derived from Human Placenta Tissue., World J Stem Cells 4, 53-61, 2012 |
Deoxyribonuclease I: 0.01% | |||||
人类 | 人类, 男性 40-60 years | Adipose derived stem cells | Collagenase: 0.25% | PBS | Blasi, A., Martino, C., Balducci, L., Saldarelli, M., Soleti, A., Navone, S., Canzi, L., Cristini, S., Invernici, G., Parati, E. and Alessandri, G.: Dermal Fibroblasts Display Similar Phenotypic and Differentiation Capacity to Fat-Derived Mesenchymal Stem Cells, but Differ in Anti-Inflammatory and Angiogenic Potential, Vasc Cell 3, 5, 2011 |
Deoxyribonuclease I: 0.002% | |||||
人类 | 人类 | Umbilical cord stromal stem | Collagenase Type 4: 0.08% | DMEM | Farias, V., Linares-Fernandez, J., Penalver, J., Paya Colmenero, J., Ferron, G., Duran, E., Fernandez, R., Olivares, E., O'Valle, F., Puertas, A., Oliver, F. and Ruiz de Almodovar, J.:Human Umbilical Cord Stromal Stem Cell Express CD10 and Exert Contractile Properties.,Placenta 32, 86, 2011 |
Neutral Protease: 0.138% | |||||
Hyaluronidase: 0.02% | |||||
人类 | 人类 | Vascular wall-resident multipotent stem cells | Collagenase Type 2: 0.2% | see reference | Klein, D., Weisshardt, P., Kleff, V., Jastrow, H., Jakob, H., and Ergun, S.: Vascular Wall-Resident CD44+ Multipotent Stem Cells Vive Rise to Pericytes and Smooth Muscle Cells and Contribute to New Vessel Maturation., PLoS ONE 6, e20540, 2011 |
Elastase: 5 u/ml | |||||
人类 | 人类 | Adipose derived stem | Collagenase Type 1: 0.1% | DMEM/Ham's F-12 | Yu, G. Floyd, ZE., Wu, X., Halvorsen, Y. and Gimble, J.: Isolation of Human Adipose-Derived Stem Cells from Lipoaspirates., Methods Mol Biol 702, 17-27, 2011 |
人类 | 人类 | Glioma stem cells | PDS kit: per instructions | neurobasal medium | Hjelmeland, A., Wu, Q., Wickman, S., Eyler, C., Heddleston, J., Shi, Q., Lathia, J., Macswords, J., Lee, J., McLendon, R. and Rich, J.: Targeting A20 Decreases Glioma Stem Cell Survival and Tumor Growth., PLoS Biol 8, e1000319, 2010 |
人类 | 人类 | Dental pulp and apical papilla stem cells | Collagenase Type 1: 0.3% | MEM | Huang, G., Yamaza, T., Shea, L., Djouad, F., Kuhn, N., Tuan, R. and Shi, S.:Stem/Progenitor Cell-Mediated De Novo Regeneration of Dental Pulp with Newly Deposited Continuous Layer of Dentin in an In Vivo Model., Tissue Eng Part A 16, 605, 2010 |
Neutral Protease: 0.4% | |||||
人类 | 人类 | Corneal stromal stem | Neutral Protease: 1.2 u/ml | DMEM | Du, Y., Roh, D., Funderburgh, M., Mann, M., Marra, K., Rubin, J., Li, X. and Funderburgh, J.: Adipose-Derived Stem Cells Differentiate to Keratocytes In Vitro., Mol Vis 16, 2680, 2010 |
Collagenase: 0.1% | |||||
人类 | 人类 | Adipose tissue-derived stem | Collagenase Type 2: 1.0% | DMEM/F12 | Tan, H., DeFail, A., Rubin, J., Chu, C. and Marra, K.: Novel Multiarm PEG-Based Hydrogels for Tissue Engineering., J Biomed Mater Res 92, 979, 2010 |
人类 | 人类 | Umbilical cord mesenchymal | Hyaluronidase: 0.05% | DMEM | Zeddou, M., Briquet, A., Relic, B., Josse, C., Malaise, M., Gothot, A., Lechanteur, C. and Beguin, Y.: The Umbilical Cord Matrix is a Better Source of Mesenchymal Stem Cells (MSC) Than the Umbilical Cord Blood., Cell Biol Int 34, 693-701, 2010 |
Collagenase: 0.08% | |||||
人类 | 人类 | Adipose stem cells | Collagenase Type 1: 0.1% | DMEM | Hareendran, S., Sathishkumar, S., Abbas, S., Mackay, A. and Rajan, P.: A Novel Composition for the Culture of Human Adipose Stem Cells which Includes Complement C3.,Cytotechnology 62, 389-402, 2010 |
人类 | 人类, 40-65 year | Adult human adipose stem cells | Collagenase Type 2: 0.075% | DMEM | Sun, N., Panetta, N., Gupta, D., Wilson, K., Lee, A., Jia, F., Hu, S., Cherry, A., Robbins, R., Longaker, M. and Wu, J.: Feeder-Free Derivation of Induced Pluripotent Stem Cells from Adult Human Adipose Stem Cells., Proc Natl Acad Sci U S A 106, 15720, 2009 |
人类 | 人类 | Filum terminale neural progenitor | Trypsin: see reference | DMEM/F-12 | Varghese, M., Olstorn, H., Berg-Johnsen, J., Moe, M., Murrell, W. and Langmoen, I.:Isolation of Human Multipotent Neural Progenitors from Adult Filum Terminale., Stem Cells Dev 18, 603, 2009 |
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, 男性 | Spermatogonial stem cells | Collagenase: 1% | DMEM | Kossack, N., Meneses, J., Shefi, S., Nguyen, H., Chavez, S., Nicholas, C., Gromoll, J., Turek, P. and Reijo-Pera, R.: Isolation and Characterization of Pluripotent Human Spermatogonial Stem Cell-Derived Cells., Stem Cells , , 2008 |
Deoxyribonuclease I: 0.22% | |||||
Trypsin: 0.4% | |||||
人类 | 人类 | Muscle derived multiprogenitor cells | Collagenase Type 2: 0.05% | DMEM | Nesti, L., Jackson, W., Shanti, R., Koehler, S., Aragon, A., Bailey, J., Sracic, M., Freedman, B., Giuliani, J. and Tuan, R.: Differentiation Potential of Multipotent Progenitor Cells Derived from War-Traumatized Muscle Tissue., J Bone Joint Surg Am 90, 2390, 2008 |
人类 | 人类 | Adipose derived stem cells | Collagenase Type 1: 0.075% | DMEM | Jeong, J.: Adipose Stem Cells as a Clinically Available and Effective Source of Adult Stem Cell Therapy, Int J Stem Cells 1, 43, 2008 |
人类 | 人类 | Hepatic stem cells and heptoblasts | Collagenase Type 4: 0.014-0.06% | various | Wauthier, E., Schmelzer, E., Turner, W., Zhang, L., LeCluyse, E., Ruiz, J., Turner, R., Furth, M., Kubota, H., Lozoya, O., Barbier, C., McClelland, R., Yao, H., Moss, N., Bruce, A., Ludlow, J. and Reid, L.: Hepatic Stem Cells and Hepatoblasts: Identification, Isolation, and Ex Vivo Maintenance., Methods Cell Biol 86, 137, 2008 |
人类 | 人类 | Umbilical mesenchymal stem | Collagenase: 0.1% | DMEM | Secco, M., Zucconi, E., Vieira, N., Fogaca, L.,Cerqueira, A., Carvalho, M., Jazedje, T., Okamoto, O., Muotri, A. and Zatz, M.: Multipotent Stem Cells from Umbilical Cord: Cord is Richer Than Blood!, Stem Cells 26, 146, 2008 |
人类 | 人类 | Stem Cells | Collagenase: 280 u/ml | D-PBS | Pilgaard, L., Lund, P., Rasmussen, J., Fink, T. and Zachar, V.: Comparative Analysis of Highly Defined Proteases for the Isolation of Adipose Tissue-Derived Stem Cells., Regen Med3, 705-15, 2008 |
人类 | 人类 | Bone marrow derived MSC | Trypsin: 0.05% | DMEM | Welter, J., Solchaga, L.and Penick, K.: Simplification of Aggregate Culture of Human Mesenchymal Stem Cells as a Chondrogenic Screening Assay, Biotechniques 42, 732, 2007 |
Papain: 0.0025% | |||||
人类 | 人类, 8-12 year | Tendon stem/progenitor | Collagenase Type 1: 0.3% | DMEM | Bi Yanming, Ehirchiou Driss, Kilts Tina M, Inkson Colette A, Embree Mildred C, Sonoyama Wataru, Li Li, Leet Arabella I, Seo Byoung-Moo, Zhang Li, Shi Songtao, Young Marian F:Identification of tendon stem/progenitor cells and the role of the extracellular matrix in their niche, Nat Med 13, 1219-27, 2007 |
Neutral Protease: 0.4% | |||||
人类 | 人类, 男性 | Adipose derived adult stem cells | Collagenase Type 1: 0.1% | DMEM | Lei, L., Liao, W., Sheng, P., Fu, M., He, A. and Huang, G.: Biological Character of Human Adipose-Derived Adult Stem Cells and Influence of Donor Age on Cell Replication in Culture.,Sci China C Life Sci 50, 320, 2007 |
人类 | 人类 | Pancreatic cancer stem cells | Collagenase Type 4: 200 u/ml | medium 199 | Li, C., Heidt, D., Dalerba, P., Burant, C., Zhang, L., Adsay, V., Wicha, M., Clarke, M. and Simeone, D.: Identification of Pancreatic Cancer Stem Cells., Cancer Res 67, 1030, 2007 |
人类 | 人类, 成年人 | Adipose-derived adult stem | Collagenase Type 1: 0.1% | DMEM/F-12 Ham's | Mitchell James B, McIntosh Kevin, Zvonic Sanjin, Garrett Sara, Floyd Z Elizabeth, Kloster Amy, Di Halvorsen Yuan, Storms Robert W, Goh Brian, Kilroy Gail, Wu Xiying, Gimble Jeffrey M: Immunophenotype of human adipose-derived cells: temporal changes in stromal-associated and stem cell-associated markers, Stem Cells 24, 376-85, 2006 |
人类 | 人类 | Mesenchymal stem | Collagenase Type 1: 0.075% | DMEM | Kern Susanne, Eichler Hermann, Stoeve Johannes, Kluter Harald, Bieback Karen:Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue, Stem Cells 24, 1294-301, 2006 |
人类 | 人类 | Placental mesenchymal stem | Collagenase Type 2: 270 u/ml | MEM | Portmann-Lanz CBettina, Schoeberlein Andreina, Huber Alexander, Sager Ruth, 男性k Antoine, Holzgreve Wolfgang, Surbek DanielV: Placental mesenchymal stem cells as potential autologous graft for pre- and perinatal neuroregeneration, Am J Obstet Gynecol194, 664-73, 2006 |
Neutral Protease: 2.4 u/ml | |||||
人类 | 人类 | Stromal stem cells | Collagenase Type 1: 0.2% | HBSS | Boquest Andrew C, Shahdadfar Aboulghassem, Brinchmann Jan E, Collas Philippe:Isolation of stromal stem cells from human adipose tissue, Methods Mol Biol 325, 35-46, 2006 |
人类 | 人类, 女性 | Adipose derived adult stem cells | Collagenase Type 1: 0.1% | DMEM/F-12 | Guilak, F., Lott, K., Awad, H., Cao, Q., Hicok, K., Fermor, B. and Gimble, J.: Clonal Analysis of the Differentiation Potential of Human Adipose-Derived Adult Stem Cells., J Cell Physiol206, 229, 2006 |
人类 | 人类 | Adult stem cells | Collagenase Type 1: 0.1% | PBS | Devireddy, R., Thirumala, S. and Gimble, J.: Cellular Response of Adipose Derived Passage-4 Adult Stem Cells to Freezing Stress., J Biomech Eng 127, 1081, 2005 |
人类 | 人类, 女性 | Adipose derived adult stem cells | Collagenase Type 1: 0.1% | DMEM-Ham's F-12 | Aust, L., Devlin, B., Foster, S., Halvorsen, Y., Hicok, K., du Laney, T., Sen, A., Willingmyre, G. and Gimble, J.: Yield of Human Adipose-Derived Adult Stem Cells from Liposuction Aspirates., Cytometry 6, 7-14, 2004 |
人类 | 人类, 成年人, 男性 | Human epidermal keratinocyte stem cells | Neutral Protease: 0.5% | DMEM | Papini S, Cecchetti D, Campani D, Fitzgerald W, Grivel J, Chen S, Margolis L, Revolla R.: Isolation and Clonal Analysis of Human Epidermal Keratinocyte Stem Cells in Long-term Culture, Stem Cells 21, 481, 2003 |
人类 | 人类 | Umbilical vein mesenchymal stem cells | Collagenase: 1% | PBS | Covas DT, Siufi JL C, Silva AR L, Orellana MD: Isolation and culture of umbilical vein mesenchymal stem cells, Braz J Med Biol Res 36, 1179-83, 2003 |
人类 | 人类, 7-8 岁 | Stem cells Human exfoliated deciduous teeth | Collagenase Type 1: 0.3% | see reference | Miura, M., Gronthos, S, Zhao, M, Lu, B, Fisher, L, Robey, P, and Shi, S.: SHED: Stem Cells from Human Exfoliated Deciduous Teeth, Proc Natl Acad Sci U S A 100, 5807, 2003 |
Neutral Protease: 0.4% | |||||
人类 | 人类, 胎儿 | Epithelial progenitor | Collagenase: 0.03% | DMEM | Malhi, H., Irani, A., Gagandeep, S. and Gupta, S.: Isolation of Human Progenitor Liver Epithelial Cells with Extensive Replication Capacity and Differentiation into Mature Hepatocytes., J Cell Sci 115, 2679, 2002 |
人类 | 人类, 成年人 | Human skin mast cells | Collagenase Type 2: 0.15% | HBSS | Kambe, N, Kambe, M, Kochan, J, and Schwartz ,L.: Human Skin-derived Mast Cells Can Proliferate While Retaining Their Characteristic Functional and Protease Phenotypes, Blood 97, 2045, 2001 |
Hyaluronidase: 0.07% | |||||
Deoxyribonuclease I: 0.03% | |||||
人类 | 人类 | Central nervous system stem | Collagenase: 0.1% | HBSS | Uchida, N., Buck, D., He, D., Reitsma, M., Masek, M., Phan, T., Tsukamoto, A., Gage, F. and Weissman, I.: Direct Isolation of Human Central Nervous System Stem Cells., Proc Natl Acad Sci U S A 97, 14720, 2000 |
Hyaluronidase: 0.1% | |||||
人类 | 人类 | Embryonic stem | Neutral Protease: 1% | DMEM | Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM: Embryonic stem cell lines derived from human blastocysts, Science 282, 1145-7, 1998 |
Collagenase Type 4: 0.1% | |||||
人类 | 人类 | Muscle-derived stem cells | Trypsin: 0.25% | DMEM/F12 | Alessandri Giulio, Pagano Stefano, Bez Alessandra, Benetti Anna, Pozzi Stefano, Iannolo Gioacchin, Baronio Manuela, Invernici Gloria, Caruso Arnaldo, Muneretto Claudio, Bisleri Gianluigi, Parati Eugenio: Isolation and culture of human muscle-derived stem cells able to differentiate into myogenic and neurogenic cell lineages, Lancet 364, 1872-83, |
猴 | 猴 | Embryonic stem | Collagenase Type 4: 0.08% | DMEM | Chen, S., Revolla, R., Papini, S., Michelini, M., Fitzgerald, W., Zimmerberg, J.,and Margolis, L.: Multilineage Differentiation of Rhesus Monkey Embryonic Stem cells in Three-dimensional Culture Systems, Stem Cells 21(3), 281, 2003 |
猴 | Baboons, 1岁, 7岁, 14 岁 | Primate spermatogonial | Collagenase Type 2: 0.1% | DMEM | Nagano, M., McCarrey, J., and Brinster, R.: Primate Spermatogonial Stem Cells Colonize Mouse Testes, Biol Reprod 64, 1409, 2001 |
Trypsin: 0.05% | |||||
Deoxyribonuclease I: 0.1% | |||||
小鼠 | 小鼠, 3-4 周 | Cardiac progenitor | Collagenase Type 1: 0.1% | DMEM | Wang, H., Chen, H., Feng, B., Wang, X., He, X., Hu, R., Yin, M., Wang, W., Fu, W. and Xu, Z.: Isolation and Characterization of a Sca-1+/CD31- Progenitor Cell Lineage Derived From Mouse Heart Tissue., BMC Biotechnol 14, 75, 2014 |
小鼠 | 小鼠, P4 | Enteric neural crest progenitors | Neutral Protease: 0.5% | DMEM/F12 | Hotta, R., Stamp, L., Foong, J., McConnell, S., Bergner, A., Anderson, R., Enomoto, H., Newgreen, D., Obermayr, F., Furness, J. and Young, H.: Transplanted Progenitors Generate Functional Enteric Neurons in the Postnatal Colon., J Clin Invest 123, 1182, 2013 |
Collagenase animal free: 0.05% | |||||
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, 10 周 | Synovial mesenchymal | Collagenase: 0.1% | DMEM | Futami, I., Ishijima, M., Kaneko, H., Tsuji, K., Ichikawa-Tomikawa, N., Sadatsuki, R., Muneta, T., Arikawa-Hirasawa, E., Sekiya, I. and Kaneko, K.: Isolation and Characterization of Multipotential Mesenchymal Cells from the Mouse Synovium., PLoS ONE7, e45517, 2012 |
Deoxyribonuclease I: 0.005% | |||||
小鼠 | 小鼠, 6-8 周 | Neural stem cells | Papain: 0.01% | DMEM/F12 | Bracko, O., Singer, T., Aigner, S., Knobloch, M., Winner, B., Ray, J., Clemenson, G., Suh, H., Couillard-Despres, S., Aigner, L., Gage, F. and Jessberger, S.: Gene Expression Profiling of Neural Stem Cells and their Neuronal Progeny Reveals IGF2 as a Regulator of Adult Hippocampal Neurogenesis., J Neurosci 32, 3376-87, 2012 |
Neutral Protease: 0.1% | |||||
Deoxyribonuclease I: 0.01% | |||||
小鼠 | 小鼠, 8-10 周 | Adipose derived stem | Collagenase Type 1: 0.025% | HBSS | Sugii, S., Kida, Y., Berggren, W. and Evans, R.: Feeder-Dependent and Feeder-Independent iPS Cell Derivation from Human and Mouse Adipose Stem Cells., Nat Protoc 6, 346, 2011 |
小鼠 | 小鼠 | Spleen, bone marrow endothelial | Collagenase Type 4: 0.3-1.0% | PBS | Shi, C., Jia, T., Mendez-Ferrer, S., Hohl, T., Serbina, N., Lipuma, L., Leiner, I., Li, M., Frenette, P. and Pamer, E.: Bone Marrow Mesenchymal Stem and Progenitor Cells Induce Monocyte Emigration in Response to Circulating Toll-Like Receptor Ligands., Immunity 34, 590, 2011 |
Deoxyribonuclease I: 20 u/ml | |||||
小鼠 | 小鼠, 8-10 周 | Lung mesenchymal stem | Collagenase Type 2: 0.2% | HBSS | Chow, K., Jun, D., Helm, K., Wagner, D. and Majka, S.: Isolation & Characterization of Hoechst(low) CD45(negative) Mouse Lung Mesenchymal Stem Cells., J Vis Exp 56, e3159, 2011 |
小鼠 | 小鼠, 新生的 | Dental Pulp Stem Cells | Collagenase Type 4: 0.2% | PBS | Janebodin, K., Horst, O., Ieronimakis, N., Balasundaram, G., Reesukumal, K., Pratumvinit, B. and Reyes, M.: Isolation and Characterization of Neural Crest-Derived Stem Cells from Dental Pulp of Neonatal Mice., PLoS ONE 6, e27526, 2011 |
Neutral Protease: 1.2 u/ml | |||||
小鼠 | 小鼠 | Stem and progenitor | Collagenase Type 2: 0.2% | HBSS | Han, J., Koh, Y., Moon, H., Ryoo, H., Cho, C., Kim, I. and Koh, G.: Adipose Tissue is an Extramedullary Reservoir for Functional Hematopoietic Stem and Progenitor Cells., Blood 115, 957, 2010 |
小鼠 | 小鼠, 6-8 周 | Bone marrow mesenchymal stem | Collagenase Type 1: 0.25% | RPMI 1640 | Xu, S., De Becker, A., Van Camp, B., Vanderkerken, K. and Van Riet, I.: An Improved Harvest and In Vitro Expansion Protocol for Murine Bone Marrow-Derived Mesenchymal Stem Cells., J Biomed Biotechnol 2010, 105940, 2010 |
小鼠 | 小鼠, 成年人 | Adult neural stem | Trypsin: 0.05% | DMEM | Deleyrolle, L. and Reynolds, B.: Isolation, Expansion, and Differentiation of Adult Mammalian Neural Stem and Progenitor Cells using the Neurosphere Assay., Methods Mol Biol 549, 91, 2009 |
小鼠 | 小鼠, 6-8 周 | Prostate epithelial/stem | Collagenase Type 2: 0.5% | HBSS | Burger, P., Gupta, R., Xiong, X., Ontiveros, C., Salm, S., Moscali, D. and Wilson, E.:High Aldehyde Dehydrogenase Activity: A Novel Functional Marker of Murine Prostate Stem/Progenitor Cells., Stem Cells 27, 2220-8, 2009 |
Trypsin: 0.05% | |||||
小鼠 | 小鼠 | Adipose tissue-derived stem | Collagenase Type 1: 0.075% | Modified Eagles | Cho, K., Park, H., Park, H., Jung, J., Jeon, S., Kim, Y. and Roh, H.: IFATS Collection: Immunomodulatory Effects of Adipose Tissue-Derived Stem Cells in an Allergic Rhinitis Mouse Model., Stem Cells 27, 259-65, 2009 |
小鼠 | 小鼠 | Germ cells | Collagenase Type 1: 100 u/ml | HBSS | Breault, D., Min, I., Carlone, D., Farilla, L., Ambruzs, D., Henderson, D., Algra, S., Montgomery, R., Wagers, A. and Hole, N.: Generation of mTert-GFP Mice as a Model to Identify and Study Tissue Progenitor Cells., Proc Natl Acad Sci U S A 105, 10420, 2008 |
小鼠 | 小鼠 | Neural progenitor cell | Papain: 10 ul/ml | PBS | Hutton, S. and Pevny, L.: Isolation, Culture, and Differentiation of Progenitor Cells from the Central Nervous System., Cold Spring Harb. Protoc. 11, 5077, 2008 |
小鼠 | 小鼠, 3-6 周 | Ear mesenchymal stem | Collagenase Type 1: 0.2% | DMEM/F12 | Staszkiewicz, J., Gimble, J., Manuel, J. and Gawronska-Kozak, B.: IFATS Collection: Stem Cell Antigen-1-Positive Ear Mesenchymal Stem Cells Display Enhanced Adipogenic Potential.,Stem Cells 26, 2666, 2008 |
小鼠 | 小鼠, 胚胎 | HES-BC cells | Trypsin: 0.05% | DMEM | Lu Shi-Jiang, Feng Qiang, Caballero Sergio, Chen Yu, Moore Malcolm A, Grant Maria B, Lanza Robert: Generation of functional hemangioblasts from human embryonic stem cells,Nat Methods 4, 501-9, 2007 |
小鼠 | 小鼠, 6-8 周 | Tendon stem/progenitor | Collagenase Type 1: 0.3% | DMEM | Bi Yanming, Ehirchiou Driss, Kilts Tina M, Inkson Colette A, Embree Mildred C, Sonoyama Wataru, Li Li, Leet Arabella I, Seo Byoung-Moo, Zhang Li, Shi Songtao, Young Marian F:Identification of tendon stem/progenitor cells and the role of the extracellular matrix in their niche, Nat Med 13, 1219-27, 2007 |
Neutral Protease: 0.4% | |||||
小鼠 | 小鼠 | Liver epithelial progenitor cells | Collagenase Type 4: 0.1% | DMEM | Li Wen-Lin, Su Juan, Yao Yu-Cheng, Tao Xin-Rong, Yan Yong-Bi, Yu Hong-Yu, Wang Xin-Min, Li Jian-Xiu, Yang Yong-Ji, Lau Joseph T Y, Hu Yi-Ping: Isolation and characterization of bipotent liver progenitor cells from adult mouse, Stem Cells 24, 322-32, 2006 |
Deoxyribonuclease I: 0.05% | |||||
小鼠 | 小鼠, 6 周 | Adipose mesenchymal stem | Collagenase: 0.2% | PBS | Di Rocco Giuliana, Iachininoto Maria Grazia, Tritarelli Alessandra, Straino Stefania, Zacheo Antonella, Germani Antonia, Crea Filippo, Capogrossi Maurizio C: Myogenic potential of adipose-tissue-derived cells, J Cell Sci 119, 2945-52, 2006 |
小鼠 | 小鼠 | Neural stem cells | Papain: see reference | DMEM/F-12 | Meletis, K., Wirta, V., Hede, S., Nister, M., Lundeberg, J. abd Frisen, J.: p53 Suppresses the Self-Renewal of Adult Neural Stem Cells., Development 133, 363, 2006 |
小鼠 | 小鼠, 7天 | Cerebellar stem cells | Papain: 10u/ml | Dulbecco's PBS | Lee, A, Kessler, J, Read, T, Kaiser, C, Corbeil, D, Huttner ,W, Johnson, J, Wechsler-Reya, R.: Isolation of Neural Stem Cells from the Postnatal Cerebellum, Nat Neurosci 8, 723, 2005 |
Deoxyribonuclease I: 250 u/ml | |||||
小鼠 | 小鼠, 胚胎 | Embryonic fibroblast feeder cells | Collagenase Type 4: 0.1% | DMEM | Schatten Gerald, Smith Joseph, Navara Christopher, Park Jong-Hyuk, Pedersen Roger:Culture of human embryonic stem cells, Nat Methods 2, 455-63, 2005 |
小鼠 | 小鼠, 1天 | Neural progenitor | PDS kit: per instructions | see reference | Seaberg, R., Smukler, S. and Van der Kooy, D.: Intrinsic Differences Distinguish Transiently Neurogenic Progenitors from Neural Stem Cells in the Early Postnatal Brain., Dev Biol 278, 71, 2005 |
小鼠 | 小鼠, 8-10 周 | Liver derived stem cells | Collagenase Type 1: 0.1% | HBSS | Kotton, D., Fabian, A. and Mulligan, R.: A Novel Stem-Cell Population in Adult Liver with Potent Hematopoietic-Reconstitution Activity., Blood 106, 1574, 2005 |
Neutral Protease: 2.4 u/ml | |||||
小鼠 | 小鼠 | Bone marrow | Collagenase Type 1: 0.15% | PBS | Bertoncello, I. and Williams, B.: Hematopoietic Stem Cell Characterization by Hoechst 33342 and Rhodamine 123 Staining., Methods Mol Biol 263, 181, 2004 |
Neutral Protease: 0.15% | |||||
小鼠 | 小鼠, 4-7 天 | Pluripotent stem cells | Collagenase Type 1: 220 u/ml | MEM | Howell, J., Lee, W., Morrison, P., Zhong, J., Yoder, M. and Srour, E.: Pluripotent Stem Cells Identified in Multiple Murine Tissues., Ann N Y Acad Sci 996, 158, 2003 |
Neutral Protease: 33 u/ml | |||||
小鼠 | 小鼠, 15 天 或 3 个月 | Neural stem cells | Papain: 0.1% | DMEM/F-12 | Gritti, A, Bonfanti, L, Doetsch, F, Caille, I, Alvarez-Buylla, A, Lim, D, Galli, R, Verdugo J, Herrera, D, and Vescovi A.: Multipotent Neural Stem Cells Reside into the Rostral Extension and Olfactory Bulb of Adult Rodents, J Neurosci 22(2), 437, 2002 |
小鼠 | 小鼠, 胚胎 | Cortical progenitors | PDS kit: per instructions | Serum free medium | Estivill-Torrus, G., Pearson, H., Van Heyningen, V., Price, D. and Rashbass, P.: Pax6 is Required to Regulate the Cell Cycle and the Rate of Progression from Symmetrical to Asymmetrical Division in Mammalian Cortical Progenitors., Development 129, 455, 2002 |
小鼠 | 小鼠, 4-8 个月 | Neural subventricular zone | Trypsin: 0.13% | DMEM/F12 | Gritti A, Frolichsthal-Schoeller P, Galli R, Parati E, Cova L, Pagano S, Bjornson C, and Vescovi A.: Epidermal and Fibroblast Growth Factors Behave as Mitogenic Regulators for a Single Multipotent Stem Cell-like Population from the Subventricular Region of the Adult Mouse forebrain, J Neurosci 19(9), 3287, 1999 |
Hyaluronidase: 0.067% | |||||
小鼠 | 小鼠, 6 周 | Muscle hematopoietic stem cells | Collagenase: 0.2% | DMEM | Jackson, K., and Goodell, M.: Hematopoietic Potential of Stem Cells Isolated from Murine Skeletal Muscle, Proc Natl Acad Sci U S A 96, 14482, 1999 |
Trypsin: 0.1% | |||||
猪 | 猪, 3-4 个月 | 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,雌性, 小于一岁 | Adipose mesenchymal stem | Collagenase Type 1: 0.1% | DMEM | Williams, K., Picou, A., Kish, S., Giraldo, A., Godke, R. and Bondioli, K: Isolation and Characterization of Porcine Adipose Tissue-Derived Adult Stem Cells., Cells Tissues Organs188, 251, 2008 |
兔 | 兔, 新西兰, 8-10 周 | Tenocytes and tendon stem cells | Collagenase Type 1: 0.3% | DMEM | Zhang, J. and Wang, J.: Characterization of Differential Properties of Rabbit Tendon Stem Cells and Tenocytes., BMC Musculoskelet Disord 11, 10, 2010 |
Neutral Protease: 0.4% | |||||
大鼠 | 大鼠 | Mesenchymal Stem Cells | Collagenase Type 1: 0.075% | DMEM | Emre, E, Yuksel, N, Duruksu, G, Pirhan, D, Subasi, C, Erman, G and Karaoz, E:Neuroprotective Effects of Intravitreally Transplanted Adipose Tissue and Bone Marrow-Derived Mesenchymal Stem Cells in an Experimental Ocular Hypertension Model.,Cytotherapy 17, 543-59, 2015 |
大鼠 | SD大鼠, 5 个月 | Mesenchymal Stromal Cells | Collagenase Type 2: 0.075% | DMEM | Veronesi, F, Torricelli, P, Della, B, Pagani, S and Fini, M: In Vitro Mutual Interaction Between Tenocytes and Adipose-Derived Mesenchymal Stromal Cells., Cytotherapy 17, 215-23, 2015 |
大鼠 | 大鼠 | Adipose Derived Stem | Collagenase Type 1: 0.1% | DMEM | Wei, X., Du, Z., Zhao, L., Feng, D., Wei, G., He, Y., Tan, J., Lee, W., Hampel, H., Dodel, R., Johnstone, B., March, K., Farlow, M. and Du, Y.: IFATS Collection: The Conditioned Media of Adipose Stromal Cells Protect Against Hypoxia-Ischemia-Induced Brain Damage In Neonatal Rats., Stem Cells 27, 478, 2009 |
大鼠 | SD大鼠, 新生 | Neutral stem cells, Schwann cells | Trypsin: 0.25% | DMEM/F12 | Zeng Yuan-Shan, Ding Ying, Wu Li-Zhi, Guo Jia-Song, Li Hai-Biao, Wong Wai-Man, Wu Wu-Tian: Co-transplantation of schwann cells promotes the survival and differentiation of neural stem cells transplanted into the injured spinal cord, Dev Neurosci 27, 20-6, 2005 |
Collagenase: 0.16% | |||||
大鼠 | 大鼠, 雄性, 200-250 克 | Neural stem cells | Papain: 0.09% | EBSS | Gobbel GT, Choi SJ, Beier S, Niranjan A: Long-term c*tion of multipotential neural stem cells from adult rat subependyma, Brain Res 980, 221, 2003 |
Deoxyribonuclease I: 0.1% | |||||
大鼠 | 大鼠, 出生后的, 成年的 | Gut Neural Crest Stem Cells | Collagenase Type 4: 0.1% | HBSS | Kruger, G., Mosher, J., Bixby, S., Joseph, N., Iwashita, T. and Morrison, S.: Neural Crest Stem Cells Persist in the Adult Gut but Undergo Changes in Self-Renewal, Neuronal Subtype Potential, and Factor Responsiveness., Neuron 35, 657, 2002 |
Trypsin: 0.025% | |||||
大鼠 | 大鼠, 成年的 | Neural stem cells | Papain: 2.5 u/ml | DMEM/F-12 | Palmer, T., Markakis, E., Willhoite, A., Safar, F., and Gage, F.: Fibroblast Growth Factor-2 Activates a Latent Neurogenic Program in Neural Stem Cells from Diverse Regions of the Adult CNS, J Neurosci 19, 8487, 1999 |
Deoxyribonuclease I: 250 u/ml | |||||
Neutral Protease: 1 u/ml | |||||
大鼠 | 大鼠 | Central nervous system stem cells | Papain: 0.01% | DMEM/F-12 | Ray, J. and Gage, F.: Neural Stem Cell Isolation, Characterization and Transplantation,Modern Techniques in Neuroscience Research,Johansson, H. and Windhorst, U., Springer-Verlag New York, 339, 1999 |
Neutral Protease: 0.1% | |||||
大鼠 | 大鼠, 胚胎 | Neurons and progenitor | Papain: 20 u/ml | EBSS | Maric, D., Maric, I. and Barker, J.: Buoyant Density Gradient Fractionation and Flow Cytometric Analysis of Embryonic Rat Cortical Neurons and Progenitor Cells., Methods 16, 247, 1998 |
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