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Feeder-free generation and transcriptome characterization of functional mesenchymal stromal cells from human pluripotent stem cells.
Stem Cell Research ( IF 1.2 ) Pub Date : 2020-09-11 , DOI: 10.1016/j.scr.2020.101990
Lidan Luo 1 , Yan Zhou 2 , Chenxi Zhang 3 , Jinrong Huang 4 , Jie Du 5 , Jinqi Liao 5 , Natasja Leth Bergholt 6 , Cody Bünger 6 , Fengping Xu 3 , Lin Lin 7 , Guangdong Tong 8 , Guangqian Zhou 9 , Yonglun Luo 10
Affiliation  

Induced mesenchymal stromal cells (iMSCs) derived from human pluripotent stem cells (PSCs) are attractive cells for regenerative medicine. However, the transcriptome of iMSCs and signature genes that can distinguish MSCs from fibroblasts and other cell types are rarely explored. In this study, we reported an optimized feeder-free method for the generation of iMSCs from human pluripotent stem cells. These iMSCs display a typical MSC morphology, express classic MSC markers (CD29, CD44, CD73, CD90, CD105, CD166), are negative for lymphocyte markers (CD11b, CD14, CD31, CD34, CD45, HLA-DR), and are potent for osteogenic and chondrogenic differentiation. Using genome-wide transcriptome profiling, we created an easily accessible transcriptome reference for the process of differentiating PSCs into iMSCs. The iMSC transcriptome reference revealed clear patterns in the silencing of pluripotency genes, activation of lineage commitment genes, and activation of mesenchymal genes during iMSC generation. All previously known positive and negative markers for MSCs were confirmed by our iMSC transcriptomic reference, and most importantly, gene classification and time course analysis identified 52 genes including FN1, TGFB1, TAGLN and SERPINE1, which showed significantly higher expression in MSCs (over 3 folds) than fibroblasts and other cell types. Taken together, these results provide a useful method and important resources for developing and understanding iMSCs in regenerative medicine.



中文翻译:

来自人多能干细胞的功能性间充质基质细胞的无饲养层生成和转录组表征。

源自人多能干细胞(PSC)的诱导间充质基质细胞(iMSC)是用于再生医学的诱人细胞。然而,极少探索能将MSC与成纤维细胞和其他细胞类型区分开的iMSC的转录组和特征基因。在这项研究中,我们报告了一种优化的无饲养层方法,可从人多能干细胞中产生iMSC。这些iMSC表现出典型的MSC形态,表达经典的MSC标记(CD29,CD44,CD73,CD90,CD105,CD166),淋巴细胞标记(CD11b,CD14,CD31,CD34,CD45,HLA-DR)阴性,并且有效用于成骨和软骨形成分化。使用全基因组转录组分析,我们为将PSC分化为iMSC的过程创建了易于访问的转录组参考。iMSC转录组参考揭示了在iMSC生成过程中多能性基因沉默,谱系承诺基因激活和间充质基因激活的清晰模式。我们的iMSC转录组学参考资料证实了所有先前已知的MSC阳性和阴性标记,最重要的是,基因分类和时程分析确定了52个基因,包括FN1,TGFB1,TAGLNSERPINE1在MSC中的表达明显高于成纤维细胞和其他细胞类型(超过3倍)。综上所述,这些结果为开发和了解再生医学中的iMSCs提供了有用的方法和重要资源。

更新日期:2020-09-11
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