Despite recent progress in the preparation of feeder cells for human induced pluripotent stem cells (hiPSCs), there remain issues which limit the acquisition of feeder cells in large scale. Approaches for obtaining feeder cells quickly on a large scale are in immediate need. To reach this goal, we established suspension-adhesion method (SAM) and three-dimensional (3D) suspension method (3DSM). In SAM, mouse embryonic fibroblast (MEF) growth were fully inhibited by 10 μg/ml mitomycin-C (MMC) in 0.5 hours, and the feeder cells generated display higher adherent and recovery rates as well as longer survival time compared to conventional method (CM). 3DSM, an optimized method of SAM in which MEFs were cultured and MMC treated in suspension, was developed to lower the costs and workload using CELLSPIN System. The yield of feeder cells is several times the yield of SAM while the adherent and recovery rates and the capacity of supporting hiPSCs growth were not sacrificed. Hence, 3DSM is an economical and easy way to generate large-scale feeder cells for hiPSCs cultures.

中文翻译：

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用于大规模制备和应用诱导多能干细胞的高效饲养细胞制备系统。

尽管最近在制备用于人诱导的多能干细胞（hiPSC）的饲养细胞方面取得了进展，但是仍然存在限制大规模获得饲养细胞的问题。迫切需要快速大规模获得饲养细胞的方法。为了达到这个目标，我们建立了悬浮-粘附方法（SAM）和三维（3D）悬浮方法（3DSM）。在SAM中，小鼠胚胎成纤维细胞（MEF）的生长在0.5小时内被10μg/ ml丝裂霉素C（MMC）完全抑制，并且与常规方法相比，生成的饲养细胞显示出更高的粘附和恢复率以及更长的生存时间（厘米）。开发了3DSM，这是一种优化的SAM方法，其中通过培养MEF并将MMC悬浮处理，从而使用CELLSPIN系统降低了成本和工作量。饲养细胞的产量是SAM产量的几倍，而粘附和回收率以及支持hiPSCs生长的能力却没有受到损失。因此，3DSM是一种为hiPSCs培养产生大规模饲养细胞的经济而简便的方法。