Department of Mechanical Engineering, Faculty of Science & Technology, Keio University, Japan.
School of Integrated Design Engineering, Graduate School of Science & Technology, Keio University, Japan.
Mater Sci Eng C Mater Biol Appl. 2018 Nov 1;92:280-286. doi: 10.1016/j.msec.2018.06.053. Epub 2018 Jun 28.
Pluripotent stem cells (PSCs), especially induced PSCs (iPSCs), have great potential for regenerative medicine. Conventionally, PSCs are cultured and expanded efficiently on feeder cell layers or on cell-adhesive matrices. Large-scale iPSC expansion in an undifferentiated state without laborious culturing procedures and high manufacturing costs for the adhesive matrix is urgently required to integrate iPSCs into therapeutic applications. For this, feeder layers or cell-adhesive matrix coating have to be removed from the iPSC culture system. To enable feeder- and matrix coating-free culture conditions, we focused on a UV/ozone surface treatment technique for polystyrene cell culture substrates to improve PSC adhesion and proliferation. In this study, changes in the molecular structure of UV/ozone-modified polystyrene were characterized to optimize the surface chemistry for iPSC. Mouse iPSCs (miPSCs) were cultured on the UV/ozone-modified polystyrene substrates without feeder layers. As a result, large polymeric chains of polystyrene were dissociated into small polymeric chains and oxidized to form ester and carboxylic acid functional groups by the UV/ozone treatment. Moreover, it was suggested that optimal valance of these modified molecules enabled the feeder- and matrix coating-free culture of miPSC with maintaining pluripotency.
多能干细胞（PSCs），特别是诱导多能干细胞（iPSCs），在再生医学方面具有巨大的潜力。传统上，PSCs 在饲养细胞层或细胞黏附基质上被高效地培养和扩增。为了将 iPSCs 整合到治疗应用中，迫切需要在无分化状态下大规模扩增 iPSCs，而无需费力的培养程序和高制造成本的黏附基质。为此，必须从 iPSC 培养系统中去除饲养层或细胞黏附基质涂层。为了实现无饲养层和无基质涂层的培养条件，我们专注于使用 UV/臭氧表面处理技术对聚苯乙烯细胞培养基板进行处理，以改善 PSC 的黏附和增殖。在这项研究中，我们对 UV/臭氧改性聚苯乙烯的分子结构变化进行了表征，以优化 iPSC 的表面化学性质。在没有饲养层的情况下，将小鼠 iPSCs（miPSCs）培养在 UV/臭氧改性的聚苯乙烯基板上。结果表明，UV/臭氧处理将大的聚苯乙烯长链解离成小的长链，并氧化形成酯和羧酸官能团。此外，这些改性分子的最佳价态有助于在不使用饲养层和基质涂层的情况下培养 miPSC 并维持其多能性。
