Ibuki Masato, Horiguchi Ikki, Sakai Yasuyuki
Regenerative Medicine and Cell Therapy Laboratories, Kaneka Corporation, 6-7-3, Minatojima Minamimachi, Chuo-ku, Kobe, Hyogo, 650-0047, Japan.
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka, 565-0871, Japan.
Regen Ther. 2019 Apr 25;12:74-82. doi: 10.1016/j.reth.2019.03.008. eCollection 2019 Dec 15.
Suspension culture for the increase in human induced pluripotent stem cells (hiPSCs) has been one of the major challenges. Previously, we reported that albumin-associated lipids prevented aggregation of hiPSCs, whereas, lipids responsible for this function were unclear. Here, by using cell aggregation assay, we investigated principal lipids regulated aggregation size of hiPSCs. As a result, lysophosphatidic acid (LPA) and Sphingosine-1-phosphate (S1P), known as lysophospholipids acting as a signaling molecule, were identified. These lipids regulated the aggregation size in a dose-dependent manner. Aggregates formed with these lipids kept the high-expression rates of pluripotent marker genes and had the abilities of proliferation. These studies demonstrated that LPA and S1P were useful for suspension culture for hiPSCs without affecting the growth ability and pluripotency of hiPSCs. This knowledge will lead to the development of a simple and robust method for the mass culture of hiPSCs.
增加人诱导多能干细胞（hiPSC）的悬浮培养一直是主要挑战之一。此前，我们报道白蛋白相关脂质可防止hiPSC聚集，然而，负责此功能的脂质尚不清楚。在此，通过细胞聚集试验，我们研究了调节hiPSC聚集大小的主要脂质。结果，鉴定出溶血磷脂酸（LPA）和1-磷酸鞘氨醇（S1P），它们是作为信号分子的溶血磷脂。这些脂质以剂量依赖性方式调节聚集大小。用这些脂质形成的聚集体保持多能性标记基因的高表达率，并具有增殖能力。这些研究表明，LPA和S1P可用于hiPSC的悬浮培养，而不影响hiPSC的生长能力和多能性。这一知识将推动开发一种简单而强大的hiPSC大规模培养方法。
