Kim Yoon Young, Min Harry, Kim Hoon, Choi Young Min, Liu Hung Ching, Ku Seung-Yup
1Department of Obstetrics and Gynecology, Seoul National University College of Medicine, 101 Daehak-ro, Jongno-gu, Seoul, 03080 Korea.
2Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University College of Medicine, 101 Daehak-ro, Jongno-gu, Seoul, 03080 Korea.
Tissue Eng Regen Med. 2017 Mar 15;14(2):163-169. doi: 10.1007/s13770-017-0051-4. eCollection 2017 Apr.
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.
微小RNA（miRNA）是参与基因表达转录和转录后调控的小型非编码RNA分子。miRNA在包括胚胎发育在内的细胞功能中发挥着多种作用。人类胚胎干细胞（hESC）能够自我更新，并可分化为包括心肌细胞（CM）在内的大多数细胞类型。hESC的这些特性使其被视为研究人类胚胎发育和组织特异性分化的重要模型。在本研究中，我们试图展示hESC向心肌细胞分化过程中miRNA的表达谱。为诱导分化，我们通过直接分化方法将hESC分化为CM，并对分化后的细胞进行表征。为分析miRNA的表达，我们区分了每个分化阶段（第4、8、12、16、20、24、28天）并分离了RNA。进行了miRNA特异性RT-qPCR，并评估了miR-1、-30d、-133a、-143、-145、-378a、-499a的表达谱。所有miR的表达在第8天均上调。miR-143和-145的表达在分化后期也上调。仅miR-378a的表达在分化的其他阶段恢复到未分化hESC的水平。总之，我们阐明了hESC向心肌细胞分化过程中miRNA的表达谱。我们的研究结果表明，miRNA的表达在分化过程中是阶段依赖性的，并表明向CM的分化可通过直接或间接途径由miRNA调控。
