Comparison of the characteristics of mesenchymal stem-like cells derived by integration-free induced pluripotent stem cells in different single-cell culture media under feeder-free conditions
Comparison of the characteristics of mesenchymal stem-like cells derived by integration-free induced pluripotent stem cells in different single-cell culture media under feeder-free conditions
复制标题
无饲养条件下不同单细胞培养基中免整合诱导多能干细胞衍生的间充质干细胞特性比较
DOI：
10.1007/s00795-018-0211-1
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发表时间：
2019
影响因子：
1.8
通讯作者：
Morita S
中科院分区：
文献类型：
--
作者：
Ueda M;Hashimoto Y;HondaY. Baba S;Morita S
从诱导多能干细胞 (iPSC) 生成间充质干细胞 (MSLC) 可能是获得用于自体组织工程的足够细胞的实用方法。在特定培养基和非饲养细胞中进行单细胞培养是克服胚胎培养问题的一种替代且有前景的策略；然而，人们对不同培养基如何影响 MSLC 的增殖和分化知之甚少。我们首先使用三种不同的细胞培养基，包括无饲养层条件下的单细胞培养基：mTeSR1、DEF-CS500 或 StemFit AK02N，从具有非整合附加型质粒载体（以下简称 409B2 细胞）的 iPSC 中衍生出 MSLC。四次传代后，所有 MSLC 的形态完全改变为成纤维细胞形态。表面抗原CD29、CD44、CD73、CD90在所有传代中均表达，但不表达CD34和CD45。 RUNX2 在所有三种无饲养层培养基中培养的 MSLC 中表达，而 SOX9 和 PPARγ 在仅在 DEF-CS500 中培养的 MSLC 中表达。来自单细胞培养基 DEF-CS500 的 MSLC 生长速度比其他培养基中培养的 MSLC 稍快，并表达三系分化的早期基因。总而言之，这些发现为使用单细胞培养方法生成 MSLC 提供了有价值的信息。
Generating mesenchymal stem-like cells (MSLCs) from induced pluripotent stem cells (iPSCs) can be a practical method for obtaining the sufficient cells for autologous tissue engineering. Single-cell culturing in specific medium and non-feeder cells is an alternative and promising strategy to overcome problems of embryo culture; however, little is known about how different culture media affect the proliferation and differentiation of MSLCs. We first derived MSLCs from iPSCs with non-integrating episomal plasmid vectors (hereafter 409B2 cells) using three different cell culture media, including single-cell culture medium in feeder-free condition: mTeSR1, DEF-CS500, or StemFit AK02N. The morphology of all MSLCs was completely altered to a fibroblastic morphology after four passages. Surface antigens CD29, CD44, CD73, CD90, but not CD34 and CD45, were expressed in all passages. RUNX2 was expressed in MSLCs cultured in all three feeder-free media, while SOX9 and PPARγ were expressed in MSLCs cultured in only DEF-CS500. MSLCs derived from DEF-CS500, which is a single-cell culture medium, grew at a slightly faster rate than those cultured in other media and expressed early-stage genes for tri-lineage differentiation. Taken together, these findings provide valuable information for generating MSLCs using single-cell culture methods.