Department of Life Sciences, National Chung Hsing University, Taichung, Taiwan.
Duogenic StemCells Corporation, Taichung, Taiwan.
Cell Transplant. 2023 Jan-Dec;32:9636897231198172. doi: 10.1177/09636897231198172.
Cell expansion of human pluripotent stem cells (hPSCs) commonly depends on Matrigel as a coating matrix on two-dimensional (2D) culture plates and 3D microcarriers. However, the xenogenic Matrigel requires sophisticated quality-assurance processes to meet clinical requirements. In this study, we develop an innovative coating-free medium for expanding hPSCs. The xenofree medium supports the weekend-free culture and competitive growth of hPSCs on several cell culture plastics without an additional pre-coating process. The pluripotent stemness of the expanded cells is stably sustained for more than 10 passages, featured with high pluripotent marker expressions, normal karyotyping, and differentiating capacity for three germ layers. The expression levels of some integrins are reduced, compared with those of the hPSCs on Matrigel. This medium also successfully supports the clonal expansion and induced pluripotent stem cell establishment from mitochondrial-defective MELAS (mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes) patient's peripheral blood mononuclear cells. This innovative hPSC medium provides a straightforward scale-up process for producing clinical-orientated hPSCs by excluding the conventional coating procedure.
人多能干细胞(hPSCs)的细胞扩增通常依赖于 Matrigel 作为二维(2D)培养板和 3D 微载体的涂层基质。然而,异源 Matrigel 需要复杂的质量保证过程以满足临床要求。在本研究中,我们开发了一种用于扩增 hPSC 的创新无涂层培养基。该无细胞培养基支持 hPSC 在几种细胞培养塑料上无周末培养和竞争生长,无需额外的预涂层过程。扩增细胞的多能干性稳定维持超过 10 个传代,其特征是高多能标志物表达、正常核型和三个胚层的分化能力。与 Matrigel 上的 hPSC 相比,一些整合素的表达水平降低。该培养基还成功支持了从线粒体缺陷型 MELAS(线粒体脑肌病、乳酸酸中毒和卒中样发作)患者外周血单核细胞中克隆扩增和诱导多能干细胞的建立。这种创新的 hPSC 培养基通过排除传统的涂层程序,为生产面向临床的 hPSC 提供了一种简单的放大工艺。