Wu Siqin, Johansson Jan, Hovatta Outi, Rising Anna
Division of Neurogeriatrics, Department of Neurobiology, Care Sciences and Society (NVS), Center for Alzheimer Research, Karolinska Institutet, Huddinge, 14157, Stockholm, Sweden.
Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, Box 7011, 75007, Uppsala, Sweden.
Cell Mol Life Sci. 2016 Apr;73(7):1479-88. doi: 10.1007/s00018-015-2053-5. Epub 2015 Oct 1.
Human pluripotent stem cells (hPSCs) hold great promise for applications in regenerative medicine and pharmaceutical development. Such applications require cell culture methods and reagents that are chemically defined, xeno-free, scalable, and low-cost. Herein, we describe non-mechanical passaging of hPSCs on spider silk films under chemically defined and xeno-free conditions. The cells were dissociated into single cells or small aggregates using Accutase or enzyme-free dissociation buffer and then passaged to spider silk films, where they expanded in monolayers until they covered the surface. Cells cultured over 10 passages on spider silk film remained karyotypically normal and pluripotent. In conclusion, a novel method for passaging dissociated hPSCs under conditions that are compatible with clinical applications is presented. The method is cost-efficient and may be useful for both research and clinical applications.
人多能干细胞（hPSCs）在再生医学和药物开发应用中具有巨大潜力。此类应用需要化学限定、无动物源、可扩展且低成本的细胞培养方法和试剂。在此，我们描述了在化学限定和无动物源条件下，hPSCs在蜘蛛丝膜上的非机械传代。使用Accutase或无酶解离缓冲液将细胞解离成单细胞或小聚集体，然后传代至蜘蛛丝膜上，在那里它们以单层形式生长直至覆盖表面。在蜘蛛丝膜上培养超过10代的细胞核型仍保持正常且具有多能性。总之，本文提出了一种在与临床应用兼容的条件下对解离的hPSCs进行传代的新方法。该方法具有成本效益，可能对研究和临床应用都有用。
