Role of mir-302 MicroRNA Family in Stem Cell Pluripotency and Renewal

  • Shi-Lung Lin
  • Shao-Yao Ying

Abstract Recent research in human embryonic stem (hES) cells has shown a highly promising potential in transplantation therapy. Nevertheless, it is very difficult to maintain the purity and pluripotency of these hES cells. In a fertilized egg, maternal materials naturally maintain stem cell renewal and totipotency before the 128-cell stage of embryonic development. Mouse oocytes lacking Dicer, a conserved ribonuclease required for microRNA (miRNA) biogenesis, arrest in the division phase of meiosis I, indicating that miRNAs play a critical role in oogenesis. We have observed that mir-302 familial microRNAs (mir-302s) were expressed at extremely high levels in mouse oocytes and human embryonic stem cells, and gradually decreased after cell differentiation. Therefore, we proposed that the mir-302 family is one of the key maternal materials essential for maintenance and renewal of the hES cell pluripotency. For this, we developed a Pol-II-based intronic miRNA expression system to transgenically express the mir-302s in several human epidermal and cancerous cell lines. Surprisingly, these mir-302s-transfected cells, namely miRNA-induced pluripotent stem (mirPS) cells, were shown to not only express all sorts of hES markers, such as Oct3/4, SSEA-3, SSEA-4, Sox2 and Nanog, but also have a highly demethylated genome similar to the reprogrammed genome of a fertilized egg. Microarray analyses further revealed that genome-wide gene expression patterns of these mirPS cells shared over 86% similarity with those of hES H1 and H9 cell lines. With certain molecular guidance ex vivo, these mirPS cells could differentiate into distinct tissue cell types, such as neuron-, chondrocyte-, fibroblast- and spermatogonia-like primordial cells. Based on these findings, we suggest that the function of mir-302s is able to not only maintain the hES cell renewal and pluripotency but also to reprogram differentiated cells into a hES-like pluripotent state, which may provide insights into areas of opportunity for therapeutic intervention.

Keywords

human embryonic stem (ES) cell induced pluripotent stem (iPS) cell stem cell renewal totipotency maternal materials microRNA (miRNA) mir-302 feeder-free culture epigenetic reprogramming cell differentiation 

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Copyright information

© Springer Science + Business Media B.V 2008

Authors and Affiliations

  • Shi-Lung Lin
    • 1
  • Shao-Yao Ying
    • 1
  1. 1.Department of Cell and Neurobiology, Keck School of MedicineUniversity of Southern CaliforniaLos AngelesUSA

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