原代细胞分离中,针对胸腺涉及到树突状,胸腺基质细胞、脂肪干细胞、基质细胞,上皮,胸腺等等的解离通常会使用酶,如何选择合适的酶来完成对应胸腺的解离,以及原代细胞采集的常规操作, 艾美捷针对胸腺相关的组织分类解离分种属、不同的细胞、酶组合推荐,推荐浓度、培养基以及参考文献,详细汇总指南:
本技术指南描述了标准实验室程序; 为建立细胞分离方案提供了一种合乎逻辑的实验方法; 并列出了许多组织特异性参考资料,以帮助开发有效的方法。更多其他组织的解离指南,点击查看:各组织分离指南。
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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 |
人 | Human | 树突状 / Dentritic | Collagenase Type 2: 0.1%; Deoxyribonuclease I: 0.002% | RPMI 1640 | Vandenabeele S, Hochrein H, Mavaddat N, Winkel K, Shortman K: Human thymus contains 2 distinct dendritic cell populations, Blood 97, 1733-41, 2001 |
小鼠 | Mouse | 胸腺基质细胞 / Thymic stromal | Collagenase Type 4: 0.02%; Neutral Protease: 0.02%; Deoxyribonuclease I: 0.0025% | RPMI-1640 | Rode, I. and Boehm, T.: Regenerative Capacity of Adult Cortical Thymic Epithelial Cells., Proc Natl Acad Sci U S A 109, 3463-8, 2012 |
小鼠 | Mouse | 基质细胞 / Stromal | Collagenase Type 3: 0.2%; Hyaluronidase: 0.1% | RPMI 1640 | Phillips, Joy, Brondstetter, T., English, C., Lee, H., Virts, E, and Thoman, M.: IL-7 Gene Therapy in Aging Restores Early thymopoiesis without Reversing Involution, J Immunol , 4869, 2004 |
小鼠 | Mouse | 树突状 / Dentritic | Collagenase Type 3: 0.1%; Deoxyribonuclease I: 325 u/ml | RPMI 1640 | Schiavoni, F, Mattei, F, Sestili, P, Borghi, P, Venditti, M, Morse, H, Belardelli, F, and Gabrieli, L: ICSBP is Essential for the Development of Mouse Type I Interferon-producing Cells and for the Generation and Activation of CD8a+ Dendritic Cells, J Exp Med 196, 1415, 2002 |
小鼠 | Mouse, 3-6 week | 胸腺 / Thymic | Collagenase Type 3: 100-400 u/ml | HBSS | Smith KM, Olson DC, Hirose R, Hanahan D: Pancreatic gene expression in rare cells of thymic medulla: evidence for functional contribution to T cell tolerance, Int Immunol 9, 1355-65, 1997 |
小鼠 | Mouse (BALB/c, C3H, C57BL/6), 1-28 day | 上皮,胸腺 / Epithelial, thymus | Neutral Protease: 1.5 痢/ml | DMEM | Ropke, C., van Deurs, B., and Petersen, O.: Short-term Cultivation of Murine Thymic Epithelial Cells in a Serum-Free Medium, In Vitro Cell Dev Biol 26, 671, 1990 |
小鼠 | Mouse, C3H, 16 wk old, female | 上皮 / Epithelial | Collagenase Type 3: 0.1% | DMEM | Ehmann, U., Shiurba, R., and Peterson, W.: Long-Term Proliferation of Mouse Thymic Epithelial Cells in Culture, In Vitro Cell Dev Biol 22 (12), 738, 1986 |
小鼠 | Mouse, Swiss, 6 wk | 胸腺 / Thymus | Collagenase Type 3: 150 u/ml | DMEM | Jones, K. and Pierre, R.: Analysis of Cellular Heterogeneity in Mouse Thymus Cultures, In Vitro 17, 431, 1981 |
大鼠 | Rat, postnatal | 胸腺 / Thymic | Trypsin: 0.05% | HBSS | Bonfanti, P., Claudinot, S., Amici, A., Farley, A., Blackburn, C. and Barrandon, Y.: Microenvironmental Reprogramming of Thymic Epithelial Cells to Skin Multipotent Stem Cells., Nature 466, 978, 2010 |
大鼠 | Rat | 胸腺上皮 / Thymic epithelial | Collagenase Type 3: 0.1% | DMEM | Masuda, A and Matsuyama, M: Epithelial Cell Lines From Rat Thymoma and Rat Thymus,Cell & Tissue Culture: Laboratory Procedures Vol. 2,Doyle, A., Griffiths, J., and Newell, D., John Wiley and Sons, Ltd., 21C:4.1, 1995 |
大鼠 | Rat (ACI/NMs X BUF/Mna) F1, male, 28 months ;Rat, (ACI/MNs) male, 8 weeks (also bovine, adult) | 上皮 / Epthelial | Collagenase Type 3: 0.1% | Eagle's MEM Serum-free | Masuda, A., Ohtsuka, K., and Matsuyama, M.: Establishment of Functional Epithelial Cell Lines from a Rat Thyoma and a Rat Thymus, In Vitro Cell Dev Biol 26, 713, 1990 |
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