Abstract: OBJECTIVE:To investigate effects of dehydrated icaritin (DICT) on the stemness of human urine-derived stem cells (hUSCs). METHODS:hUSCs were isolated by centrifugation at room temperature and cultured in vitro. Morphological observations of hUSCs was performed under inverted phase contrast microscope. The growth curve of hUSCs from 1-7 days was plotted according to MTT experiment. The surface markers of hUSCs were detected by flow cytometry. The control group of hUSCs culture medium and DICT treatment groups with concentrations of 0.5,1.0,1.5,2.0,2.5,3.0 μmol/L were set up. Cell viability of each group was measured by the tetramethylthiazole blue (MTT) assay from 1-7 days. Effects of 2.0 μmol/L DICT treatment of hUSCs on the expression of OCT-4 and C-MYC mRNA were measured by quantitative real-time PCR (qPCR) after 3 days. RESULTS:hUSCs with good morphology and high growth activity were successfully obtained. hUSCs expressed CD44 and CD90,surface markers of MSCs,but not CD34,surface marker of hematopoietic stem cells,and CD31,surface marker of endothelial cells. 2.0 μmol/L DICT treatment for 3-6 days significantly promoted proliferations of hUSCs,enhanced their growth viability (P<0.01),and up-regulated the mRNA expression of hUSCs stemness genes OCT-4 and C-MYC (P<0.01). CONCLUSION:DICT enhanced the stemness of hUSCs in vitro.

Key words: dehydrated icaritin, mesenchymal stem cells, human urine-derived stem cells, stemness