Abstract 515: Non-invasive Method of Generating Human Induced-mesenchymal Stem Cells Derived From Urinary Epithelial Cells for Regenerative Therapy

Abstract

Introduction: Mesenchymal stem cells (MSCs) are multipotent adult stem cells having an extensive proliferation capacity in vitro and in vivo. These MSCs can differentiate into various mesoderm-type cells such as osteoblasts, cardiomyocytes, etc. A subpopulation of urinary epithelial cells (UECs) have been identified in urine samples, is considered a promising cell resource for generating autologous induced-pluripotent stem cells (iPSCs). Hypothesis: We hypothesize that the production of high quality, autologous, induced-MSCs (iMSCs) with high replicative potential suitable for the regenerative therapy, using an easy, and the most non-invasive method of isolation, from human UECs. Methods and Results: Human urine was collected and centrifuged to obtain the UECs, which were characterized by the expression of CK19 and ZO1. These UECs were reprogrammed to iPSCs using a cocktail of mRNAs (OCT4, KLF4, SOX2, c-MYC, Nanog and Lin28) along with Lipofectamine for 11 days in culture. These iPSCs were characterized by the expression of the pluripotent markers such as OCT4, SOX2 and SSEA4. The iPSCs were subsequently differentiated into iMSCs using the mesenchymal specific medium for 21 days. iMSCs were harvested at the end of 21 days, and they were characterized by the high levels of mRNA and protein expressions of mesenchymal specific markers such as CD73, CD90 and CD105 (Fig. 1A). FACS analysis showed that more than 93% of the cells were positive for the markers of MSCs (Fig. 1B) . Moreover, the obtained iMSCs have high proliferation capacity compared with the adult stem cells. Conclusions: We have developed an easy, non-invasive method for obtaining autologous, non-immunogenic and highly-proliferating iMSCs suitable for various regenerative therapies including cardiac diseases, from urinary epithelial cells.