Suzuki Hayato, Kasai Kohei, Kimura Yuka, Miyata Shogo
School of Integrated Design Engineering, Graduate School of Science & Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Kanagawa 223-8522, Japan.
Department of Mechanical Engineering, Faculty of Science & Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Kanagawa, 223-8522, Japan.
Mater Sci Eng C Mater Biol Appl. 2021 Apr;123:112012. doi: 10.1016/j.msec.2021.112012. Epub 2021 Mar 5.
Culturing pluripotent stem cells effectively requires substrates coated with feeder cell layers or cell-adhesive matrices. It is difficult to employ pluripotent stem cells as resources for regenerative medicine due to risks of culture system contamination by animal-derived factors, or the large costs associated with the use of adhesive matrices. To enable a coating-free culture system, we focused on UV/ozone surface modification and atmospheric pressure plasma treatment for polystyrene substrates, to improve adhesion and proliferation of pluripotent stem cells. In this study, to develop a feeder- and matrix coating-free culture system for embryonic stem cells (ESCs), mouse ESCs were cultured on polystyrene substrates that were surface-modified using UV/ozone-plasma combined treatment. mESCs could be successfully cultured under feeder-free conditions upon UV/ozone-plasma combined treatment of culture substrates, without any further chemical treatments, and showed similar proliferation rates to those of cells grown on the feeder cell layer or matrix-coated substrates.
有效培养多能干细胞需要用饲养细胞层或细胞粘附基质包被的底物。由于存在动物源性因子污染培养系统的风险,或者使用粘附基质的成本高昂,因此难以将多能干细胞用作再生医学的资源。为了实现无包被的培养系统,我们专注于对聚苯乙烯底物进行紫外线/臭氧表面改性和大气压等离子体处理,以改善多能干细胞的粘附和增殖。在本研究中,为了开发一种用于胚胎干细胞(ESC)的无饲养层和无基质包被的培养系统,将小鼠胚胎干细胞培养在经紫外线/臭氧-等离子体联合处理进行表面改性的聚苯乙烯底物上。在对培养底物进行紫外线/臭氧-等离子体联合处理后,小鼠胚胎干细胞能够在无饲养层条件下成功培养,无需任何进一步的化学处理,并且其增殖速率与在饲养细胞层或基质包被底物上生长的细胞相似。