Early prediction of the differentiation potential during the formation of human iPSC-derived embryoid bodies
Early prediction of the differentiation potential during the formation of human iPSC-derived embryoid bodies
复制标题
人类 iPSC 衍生胚状体形成过程中分化潜力的早期预测
DOI：
10.1016/j.bbrc.2019.06.081
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发表时间：
2019-08-27
影响因子：
3.1
通讯作者：
Li, Yu-Mei
中科院分区：
文献类型：
--
作者：
Guo, Ning-Ning;Liu, Li-Ping;Li, Yu-Mei
诱导多能干细胞(IPSCs)即使在相同的条件下，其分化潜能也表现出巨大的差异。然而，预测这些分化趋势的方法仍然很少，特别是在分化的早期阶段。本研究旨在建立一种简单实用的拟胚体(EBS)早期参数预测IPSC系分化趋势的方法。我们比较了四个人IPSC系的EBS的形态和维持性以及它们在三个胚层的特异标记的基因表达水平。此外，还比较了这些IPSC系向外胚层来源的黑素细胞的分化潜能，并与上述参数进行了相关性分析。结果表明，形成规则的、光滑的、不囊状的EBS的IPSC系可以在培养较长时间内保持相对较长的时间，同时也表达较高水平的外胚层特异性标记和较低水平的中胚层/内胚层标记。此外，这些IPSC系在基于EB的方法中显示出更大的黑素细胞分化潜力，并且诱导的黑素细胞表达黑素细胞标志，并呈现出与正常人类黑素细胞相似的特征。相比之下，形成具有亮或暗空腔的囊状EBS并表达相对较低水平的外胚层特异性标记的IPSC系在黑素细胞分化中失败。综上所述，人IPSC系的分化趋势可以通过EB期的特定参数来预测。最适EBS的形成和维持以及胚层特异性标记的表达对于早期的预测分析尤为重要和实用。(C)2019 Elsevier Inc.保留所有权利。
Induced pluripotent stem cells (iPSCs) show huge variations in their differentiation potential, even in the same condition. However, methods for predicting these differentiation tendencies, especially in the early stage of differentiation, are still scarce. This study aimed to establish a simple and practical system to predict the differentiation tendency of iPSC lines using embryoid bodies (EBs) with identified parameters in the early stage. We compared four human iPSC lines in terms of the morphology and maintenance of EBs and their gene expression levels of specific markers for three germ-layers. Furthermore, the differentiation potentials of these iPSC lines into melanocytes, which are ectoderm-derived cells, were also compared and correlated with the above parameters. The results showed that iPSC lines forming regular, smooth, and not cystic EBs, which could be maintained in culture for a relatively longer time, also expressed higher levels of ectoderm-specific markers and lower levels of mesoderm/endoderm markers. Additionally, these iPSC lines showed greater potential in melanocyte differentiation using EB-based protocol, and the induced melanocytes expressed melanocytic markers and presented characteristics that were similar to those of normal human melanocytes. By contrast, iPSC lines that formed cystic EBs with bright or dark cavities and expressed relatively lower levels of ectoderm-specific markers failed in the melanocyte differentiation. Collectively, the differentiation tendency of human iPSC lines may be predicted by specific parameters in the EB stage. The formation and maintenance of optimal EBs and the expression of germ layer-specific markers are particularly important and practical for the prediction assay in the early stage. (C) 2019 Elsevier Inc. All rights reserved.