Howard Hughes Medical Institute, The Salk Institute for Biological Studies, La Jolla, California, USA.
Nat Protoc. 2011 Mar;6(3):346-58. doi: 10.1038/nprot.2010.199. Epub 2011 Feb 24.
Adipose tissue is an abundantly available source of proliferative and multipotent mesenchymal stem cells with promising potential for regenerative therapeutics. We previously demonstrated that both human and mouse adipose-derived stem cells (ASCs) can be reprogrammed into induced pluripotent stem cells (iPSCs) with efficiencies higher than those that have been reported for other cell types. The ASC-derived iPSCs can be generated in a feeder-independent manner, representing a unique model to study reprogramming and an important step toward establishing a safe, clinical grade of cells for therapeutic use. In this study, we provide a detailed protocol for isolation, preparation and transformation of ASCs from fat tissue into mouse iPSCs in feeder-free conditions and human iPSCs using feeder-dependent or feeder/xenobiotic-free processes. This protocol also describes how ASCs can be used as feeder cells for maintenance of other pluripotent stem cells. ASC derivation is rapid and can be completed in <1 week, with mouse and human iPS reprogramming times averaging 1.5 and 2.5 weeks, respectively.
脂肪组织是一种丰富的增殖和多能间充质干细胞来源，具有很有前途的再生治疗潜力。我们之前的研究表明，人类和小鼠脂肪来源的干细胞（ASCs）可以被重编程为诱导多能干细胞（iPSCs），其效率高于其他细胞类型的报道。ASC 衍生的 iPSCs 可以在无饲养层的情况下产生，这代表了一种研究重编程的独特模型，也是朝着建立安全的、临床级别的细胞用于治疗应用迈出的重要一步。在这项研究中，我们提供了一个详细的协议，用于从脂肪组织中分离、准备和转化 ASC 为无饲养层条件下的小鼠 iPSCs 和依赖饲养层或无饲养层/异生物质的过程中的人 iPSCs。该方案还描述了如何将 ASC 用作其他多能干细胞的饲养细胞。ASC 的衍生过程迅速，可在<1 周内完成，小鼠和人类 iPS 重编程时间平均分别为 1.5 和 2.5 周。
