Shinohara Marie, Choi Hyunjin, Ibuki Masato, Yabe Shigeharu G, Okochi Hitoshi, Miyajima Atsushi, Sakai Yasuyuki
Institute of Industrial Science, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan.
Regenerative Medicine and Cell Therapy Laboratories, Kaneka Corporation, Kobe MI R&D Center 3F, 6-7-3, Minatojima Minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.
Regen Ther. 2019 May 30;12:14-19. doi: 10.1016/j.reth.2019.05.004. eCollection 2019 Dec 15.
A differentiation of human induced pluripotent stem cells (hiPSCs) into definitive endoderm linage is required for a preparation of metabolic organ derived cells. The differentiation consumed high-priced cytokines and small molecules, which have hampered the manufacturability of differentiated cells. Although the cytokines and small molecules are remained or cells produce the autocrine factors, daily culture medium change should be proceeded to remove toxic metabolites generated from cells. In this study, we developed a simple dialysis culture system to refine the medium during definitive endodermal differentiation. We demonstrated that dialysis culture prevented cell damage to remove lactate. The hiPSCs cultured with dialysis also differentiated similarly as usual differentiation without dialysis even if they were not supplied Activin A for latter culture days in the differentiation. With this dialysis culture system, hiPSCs were differentiated into endodermal lineage with medium refinement and recycling and autocrine factors as well as cytokines, which may lead to reduce differentiation cost.
为了制备代谢器官衍生细胞，需要将人诱导多能干细胞（hiPSC）分化为确定的内胚层谱系。这种分化消耗高价细胞因子和小分子，这阻碍了分化细胞的可制造性。尽管细胞因子和小分子会残留或细胞会产生自分泌因子，但仍应每天更换培养基以去除细胞产生的有毒代谢物。在本研究中，我们开发了一种简单的透析培养系统，以在确定的内胚层分化过程中优化培养基。我们证明透析培养可防止细胞损伤以去除乳酸。即使在分化后期培养天数不供应激活素A，用透析培养的hiPSC与未进行透析的常规分化一样能正常分化。通过这种透析培养系统，hiPSC通过培养基优化、循环利用以及自分泌因子和细胞因子分化为内胚层谱系，这可能会降低分化成本。
