Abstract
We aimed to investigate the differentially expressed long non-coding RNAs (lncRNAs) during the differentiation of human umbilical cord derived mesenchymal stem cells (hUCMSCs) into cardiomyocyte-like cells induced by 5-aza. hUCMSCs were isolated and purified from umbilical cords. After treated with 10 μmol/L 5-Aza for 24 h, hUCMSCs wereas continued to be cultured for 14 days. Comparison of cardiac specific genes and the expression profile of lncRNAs on hUCMSCs between day 14 and day 0 was performed using immunofluorescence staining, immunohistochemistry, Western blot assay, RT-PCR and lncRNA microarray. Results show that well-organized sarcomeric structure and more cTnI and MLC2a staining were seen in hUCMSCs of day 14 after 5-aza-induced compared to those in day 0. Expression of Desmin, Nkx2.5, cTnI and MLC2a of hUCMSCs was much higher on day 14 compared with day 0 (P < 0.01). 41 differentially expressed lncRNAs were found on day 14 hUCMSCs compared those of day 0 were identified. Among them, 25 upregulated and 16 downregulated. Four out of the five upregulated lncRNAs (P = 0.00035, 0.014, 0.016 and 0.005 for uc010vei.1, X72487, BC064139, AK092074) and four out of the five downregulated lncRNAs (P = 0.038, 0.0014, 0.00026 and 0.004 for X85157, uc007keu.1, AK309872, NR_029399) showed significantly different expressions in further validation using RT-PCR. Our results illustrated that there was a dysregulation of the lncRNA profile during the differentiation of hUCMSCs into cardiomyocyte-like cells, which will provide the foundation for further study of the biological functions and mechanism of lncRNAs in the differentiation of hUCMSCs into cardiomyocyte-like cells.
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We are thankful Hai-hui Sheng for technical assistance and Wen-qi Li for the assistance of statistical analysis.
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ZBR carried out the design and coordinated the study, participated all the experiments and prepared the manuscript. LZ provided assistance in the design of the study, coordinated and carried out most of the experiments and participated in manuscript preparation. GCC and YR provided assistance for all experiments. All authors have read and approved the content of the manuscript.
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This study is approved by institutional ethics committee of Jiangsu Taizhou People’s Hospital and written informed consent was obtained from all patients.
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Ruan, Zb., Chen, Gc., Ren, Y. et al. Expression profile of long non-coding RNAs during the differentiation of human umbilical cord derived mesenchymal stem cells into cardiomyocyte-like cells. Cytotechnology 70, 1247–1260 (2018). https://doi.org/10.1007/s10616-018-0217-5
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DOI: https://doi.org/10.1007/s10616-018-0217-5