MicroRNAs (miRNAs) are small non-coding RNA molecules that participate in transcriptional and post-transcriptional regulation of gene expression. miRNAs have numerous roles in cellular function including embryonic development. Human embryonic stem cells (hESCs) are capable of self-renewal and can differentiate into most of cell types including cardiomyocytes (CMs). These characteristics of hESCs make them considered as an important model for studying human embryonic development and tissue specific differentiation. In this study, we tried to demonstrate the profile of miRNA expression in cardiac differentiation from hESCs. To induce differentiation, we differentiated hESCs into CMs by direct differentiation method and characterized differentiated cells. To analyze the expression of miRNAs, we distinguished (days 4, 8, 12, 16, 20, 24, 28) and isolated RNAs from each differentiation stage. miRNA specific RT-qPCR was performed and the expression profile of miR-1, -30d, -133a, -143, -145, -378a, -499a was evaluated. The expression of all miRs was up-regulated at day 8. miR-143 and -145 expression was also up-regulated at the later stage of differentiation. Only miR-378a expression returned to undifferentiated hESC levels at the other stages of differentiation. In conclusion, we elucidated the expression profile of miRNAs during differentiation into cardiomyocytes from hESCs. Our findings demonstrate the expression of miRNAs was stage-dependent during differentiation and suggest that the differentiation into CMs can be regulated by miRNAs through direct or indirect pathway.
MicroRNA Human embryonic stem cell Cardiac lineage Differentiation
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This study was supported by grants of Ministry of Science, ICT and Future Planning (2014R1A1A305264 and 2016R1E1A1A01943455). The authors would like to express sincere thanks to the technical assistance of Kyung Mee Cho and Mi Ae Lee.
Compliance with ethical standards
Conflict of interest
The authors have no conflicts of interest.
The study was approved by the IRB of the IRMP, MRC, Seoul National University (219932-201412-LR-12-01-01).
Supplementary movie. Representative beating cluster of cardiomyocytes differentiated from human embryonic stem cells (MPG 5067 kb)
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