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Worthington組織消化指南
引言
組織消化酶主要用于組織培養和細胞生物學研究的原代細胞的分離和收集。盡管這些消化酶已經普遍應用多年，但是對其組織消化和細胞收集的作用機制卻沒有全面的理解。 由此導致的結果便是選擇一種消化方法大多是隨機選擇或基于過去的經驗，而不是在*理解消化原理的基礎上選擇的消化方案，以及如何根據具體實驗目的修正消化方案以獲得更好的實驗結果。
細胞分離程序的目的是zui大程度的獲得功能活性的單個細胞，而影響結果的原因有很多，包括（不限于）以下：
1、組織類型
2、物種來源
3、動物年齡
4、遺傳改造情況（如knockouts）
5、所用的消化培養液
6、所用的酶
7、消化酶中的雜質
8、所用消化酶的工作濃度
9、溫度
10、孵育時間
通常前四個原因沒辦法選擇，而剩下的原因一般由經驗決定以獲取zui適實驗結果。
通過查閱文獻來選擇*組織消化酶和zui適消化條件的研究人員通常被互相矛盾的實驗數據搞得一頭霧水。這些巨大差異來源于組織胞外基質的復雜而動態的特性，以及過去常用的粗制、未提純的組織分離酶。另外，組織胞外基質由各種蛋白、糖蛋白、脂類和糖脂組成，這些成分的組成比例在不同物種、不同組織、甚至不同發育（年齡）之間存在明顯的差異，這些因素都極大的影響了消化方案的制定。目前通常使用的粗制酶，如鏈霉蛋白酶、NF1:250、以及含有幾種其他不同濃度蛋白酶的膠原酶、以及各種多糖酶、核酶和脂酶。
本指南總結了目前常用組織消化酶的zui適消化和細胞收集知識和方法，描述了標準的實驗步驟， 以便于制定一個*的細胞分離方案，同時列舉了一些相關的參考文獻。
下表列出多種組織的適宜消化酶（含參考文獻，以組織類型和物種分類，詳見鏈接）
脂肪組織 | 腎上腺組織 | 骨組織 | 腦組織 |
軟骨組織 | 結腸組織 | 內皮組織 | 上皮組織 |
眼 | 心臟 | 腸 | 腎 |
肝 | 肺 | 淋巴結 | 乳腺 |
其它組織 | 其他組織 | 肌組織 | 神經組織 |
胰腺 | 腮腺 | 垂體 | 前列腺 |
生殖 | 動物鱗片 | 皮膚 | 脾臟 |
干細胞 | 胸腺 | 甲狀腺/甲狀旁腺 | 扁桃體 |
腫瘤 |
|
|
|
注：使用Worthington公司消化酶的文獻不限于上表鏈接中所列，一般而言，Worthington公司的組織消化酶可用于大多組織類型的消化。
干細胞組織消化手冊
| 物種 | 詳細來源 | 細胞名稱 | 消化用酶 | 培養基 | 參考文獻 |
| 鳥 | 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 |
| 馬 | 馬 | 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 |
| 馬 | 馬, 1-5 歲 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類, 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 |
| 人類 | 人類, 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 |
| 人類 | 人類 | 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偽 Transduction.,PLoS ONE 9, e104133, 2014 |
| Trypsin: 0.25% | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類, 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類, 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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 | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類, 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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% | |||||
| 人類 | 人類, 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% | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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% | |||||
| 人類 | 人類, 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% | |||||
| 人類 | 人類, 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類, 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類 | 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 | |||||
| 人類 | 人類 | 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 |
| 人類 | 人類, 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類, 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 |
| 人類 | 人類, 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 |
| 人類 | 人類 | 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 |
| 人類 | 人類, 7-8 歲 | 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% | |||||
| 人類 | 人類, 胎兒 | 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 |
| 人類 | 人類, 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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% | |||||
| 人類 | 人類 | 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歲, 7歲, 14 歲 | 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 個月 | 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 個月 | 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% | |||||
| 豬 | 豬, 3-4 個月 | 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 |
| 豬 | 豬,雌性, 小于一歲 | 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, 新西蘭, 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 個月 | 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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