Sugimine Yoshinori, Niwa Akira, Matsubara Hiroyuki, Kobayashi Kazuki, Tabata Yasuhiko, Heike Toshio, Nakahata Tatsutoshi, Saito Megumu K
Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan.
Department of Pediatrics, Kyoto University Graduate School of Medicine, Kyoto, 6068507, Japan.
Int J Hematol. 2016 Dec;104(6):647-660. doi: 10.1007/s12185-016-2088-x. Epub 2016 Sep 6.
Various systems for differentiating hematopoietic cells from human pluripotent stem cells (PSCs) have been developed, although none have been fully optimized. In this report, we describe the development of a novel three-dimensional system for differentiating hematopoietic cells from PSCs using collagen sponges (CSs) reinforced with poly(ethylene terephthalate) fibers as a scaffold. PSCs seeded onto CSs were differentiated in a stepwise manner with appropriate cytokines under serum-free and feeder-free conditions. This process yielded several lineages of floating hematopoietic cells repeatedly for more than 1 month. On immunohistochemical staining, we detected CD34+ cells and CD45+ cells in the surface and cavities of the CS. Taking advantage of the portability of this system, we were able to culture multiple CSs together floating in medium, making it possible to harvest large numbers of hematopoietic cells repeatedly. Given these findings, we suggest that this novel three-dimensional culture system may be useful in the large-scale culture of PSC-derived hematopoietic cells.
尽管尚未完全优化，但已经开发出了多种用于从人多能干细胞（PSC）中分化造血细胞的系统。在本报告中，我们描述了一种新型三维系统的开发，该系统使用用聚对苯二甲酸乙二酯纤维增强的胶原海绵（CS）作为支架，从PSC中分化造血细胞。接种到CS上的PSC在无血清和无饲养层条件下，通过适当的细胞因子进行逐步分化。这个过程在1个多月的时间里反复产生了几类漂浮的造血细胞谱系。免疫组织化学染色显示，我们在CS的表面和腔隙中检测到了CD34+细胞和CD45+细胞。利用该系统的便携性，我们能够将多个CS一起培养在培养基中漂浮，从而有可能反复收获大量造血细胞。基于这些发现，我们认为这种新型三维培养系统可能有助于大规模培养PSC来源的造血细胞。
