Experimental study of mesenchymal stem cells derived from human umbilical cord on differatiation into functional granulosa-like cells

Abstract

Objective To investigate whether human umbilical cord mesenchymal stem cells (HUMSCs) could be induced and differentiated into granuosa cells (GCs). Methods HUMSCs were pre-treated with bone morphogenetic protein 4 (BMP4), then co-cultured with human follicular fluid and cumulus oophorus GCs. The human GCs were severed as positive control. The morphology and expression of follicle-stimulating hormone receptor (FSHR), anti-Mullerian hormone (AMH) and cytochrome P450 19A1 (CYP19A1) as well as OCT4 of cultured cells were observed and detected through immunofluorescence and fluorescent real time-PCR (RT-PCR). Results After co-culture for 12 d, the morphology of cultured HUMSCs was similar to those human GCs and FSHR positive, although intension of the fluorescence signal did not reach to that of human cumulus GCs (positive control), while HUMSCs before co-culture (negative control) showed no green fluorescence signal of FSHR. Moreover, the expression of pluripotency-related genes OCT4 in co-cultured cells down-regulated significantly (all P<0.001), and were close to that in human GCs (positive control). Meanwhile, the expression of GCs specific gene makers such as FSHR, AMH and CYP19A1 in co-cultured cells up-regulated significantly (P<0.001). The results suggested that HUMSCs had lost its stemness and differentiated into functional GCs after co-culture with human follicular fluid and cumulus oophorus GCs. Conclusion HUMSCs could be successfully induced and differentiated into functional granulosa-like cells under co-culture system of human follicular fluid and GCs. Key words: Human umbilical cord mesenchymal stem cells; Granuosa cells; Differentiation