Exosomes derived from human umbilical cord mesenchymal stem cells accelerate growth of VK2 vaginal epithelial cells through MicroRNAs in vitro
Zhu Zhongyi, Zhang Yijing, Zhang Yiqun, Zhang Hongdao, Liu Wei, Zhang Ning, Zhang Xiaodan, Zhou Guannan, Wu Ligang, Hua Keqin, Ding Jingxin
Journal:HUMAN REPRODUCTION
IF:4.99
DOI:10.1093/humrep/dey344
PMID:30576496
Published:2018-12-20
research field:分子生物学妇科干细胞生物学细胞治疗
Abstract
STUDY QUESTIONCould human umbilical cord mesenchymal stem cell-derived exosomes (hucMSC-Ex) accelerate vaginal epithelium cell (VK2) growth?SUMMARY ANSWERHucMSC-Ex play a significant role in promoting proliferation of VK2 cells by accelerating the cell cycle and inhibiting apoptosis through exosomal microRNAs in vitro.WHAT IS KNOWN ALREADYNumerous studies have reported that MSC-Ex play an important role in tissue injury repair.STUDY DESIGN, SIZE, DURATIONhucMSC and exosomes isolated from their conditioned medium were used to treat a vaginal epithelial cell line (VK2). Normal human fibroblasts (HFF-1) were used as negative control to hucMSC.PARTICIPANTS/MATERIALS, SETTING, METHODSVK2 cells were co-cultured with hucMSC whose paracrine effect on the viability, cell cycle and cell apoptosis of VK2 vaginal epithelial cells was further assessed by the CCK-8 assay and flow cytometry. HucMSC-Ex isolated from culture medium by ultracentrifuge were characterized by transmission electron microscopy, nanoparticle tracking analysis and Western blot. HucMSC-Ex at different concentrations and HFF-1 exosomes were used to treat VK2 cells. High-throughput RNA sequencing was utilized to reveal the profile of microRNAs in hucMSC, hucMSC-Ex, HFF-1 and HFF-1 exosomes and GO analysis was applied to demonstrate their functions. To evaluate the function of these specific microRNAs in hucMSC-Ex, VK2 cells were treated with RNA-interfered-hucMSC-Ex (RNAi-hucMSC-Ex) and their proliferation was measured by Label-free Real-time Cellular Analysis System.MAIN RESULTS AND THE ROLE OF CHANCEThe study showed that hucMSC stimulate VK2 cell growth possibly through a paracrine route by promoting cell cycle and inhibiting apoptosis. Compared with control and low dose groups, hucMSC-Ex of high concentration (more than 1000 ng/ml) significantly increased VK2’s growth after treatment in a dose-depended manner (P < 0.05). HucMSC-Ex raised the proportion of cells in S-phase and reduced the percentage of apop
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