Studies On The Role Of MicroRNAs In The Adipogenesis From Pocine Pluripotent Stem Cells

Terminally differentiated somatic cells can be reprogrammed into inducedpluripotent stem cells (iPSCs). It is of great significant in the research ofdevelopment mechanism and can be widely used into regenerative medicine in thefuture. As Chinese most important economic animals, pigs show great value in bothanimal genetic breeding and biomedical applications. However, there is not anyembryonic stem cells established in true sense. The appearance of iPSCs leads us anew method to further study the porcine early embryonic development.As a class of non-coding RNA, miRNAs play a major role in the regulation oftarget gene transcription level, more and more studies show that miRNAs play animportant part in regulating developmental processes in mammals. In this study, wefirst packaged the pluripotent factors Oct4, Sox2, Klf4, c-Myc retrovirus whichinfected into pocine fetal fibroblasts (PFF), and established piPSCs with ES-likecharacteristics. Then, we tried to differentiated piPSCs into adipocyte with chemicalmethod. At the same time, analyze the different expression of miRNAs in piPSCsand piPSCs-derived adipocytes with miRNA micro-array, and predicted theadipogenic differentiation pathway in this process. In order to explore if themiRNAs could affect the induction efficiency, we choose pBMSCs as donor cells,which have multiple differentiation potential and more stable adipogenicdifferentiated system, and choose miR-17, miR-21, miR-143as research target. Theresults show that:(1) the pocine fetal fibroblasts could induced into piPSCs byretrovirus-mediated OKSM four factors strategy. The piPSCs not only expressed thepluripotency factors, but also have differentiation ability into three germ layers invivo and in vitro.(2) piPSCs could differentiate into adipocyte in vitro, and theexpression of miR-17, miR-21and miR-143obviously increased in this process.(3)miR-17, miR-21and miR-143could significantly improve the efficiency ofadipogenic differentiation of pBMSCs. We constructed a stable piPSCs which not only express the multiple genes, butalso have the ability to form teratoma in NOD/SCID mice in this study, certified amore comprehensive pluripotency of piPSCs. After we obtain piPSCs, we couldhave a new way to understand the pig early embryonic development. We furtherexplore and found that piPSCs and BMSCs could differentiate into adipocytes andhave a broad application prospects in diabetes and obesity. In the process ofadipogenic differentiation of pBMSCs, we found miR-17, miR-21and miR-143could increase the adipogenic diffrentiation efficiency obviously.