The Key Laboratory of Pathobiology, Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, China.
Department of Gynecology, Third Hospital of Jilin University, Changchun, China.
Sci Rep. 2017 Sep 25;7(1):12266. doi: 10.1038/s41598-017-10428-5.
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.
尽管在制备人诱导多能干细胞（hiPSC）饲养细胞方面取得了一些进展，但仍存在一些问题限制了饲养细胞的大规模获取。因此，需要迅速大规模获得饲养细胞的方法。为了达到这个目标，我们建立了悬浮贴壁法（SAM）和三维（3D）悬浮法（3DSM）。在 SAM 中，用 10μg/ml 的丝裂霉素 C（MMC）在 0.5 小时内完全抑制了小鼠胚胎成纤维细胞（MEF）的生长，与传统方法（CM）相比，产生的饲养细胞具有更高的贴壁和恢复率以及更长的存活时间。3DSM 是 SAM 的一种优化方法，将 MEFs 在悬浮液中培养并进行 MMC 处理，以 CELLSPIN 系统降低成本和工作量。与 SAM 相比，饲养细胞的产量增加了数倍，而贴壁和恢复率以及支持 hiPSC 生长的能力并没有降低。因此，3DSM 是一种经济且简便的方法，可用于大规模生成 hiPSC 培养所需的饲养细胞。
