In vitro maturation of granulose cell by adenovirus mediated gene transduction employing follicle stimulating hormone receptor gene.

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Since the granulosa cells rapidly lose the expression of follicle stimulating hormone receptor (FSHR) and the function of steroidogenesis in primary culture conditions, co-culture of immature oocytes with granulosa cell has limitation to support oocyte maturation satisfactory for clinical application. Then, we tried to maintain the in-vitro granulosa cell function by adenoviral-mediated FSHR gene transduction. The number of FSHR on the surface of the granulosa cells prepared from the rat primed by diethylstilbestrol was 4,874 per cell immediately after sacrifice, then it decreased to 2,176 in by the third day, and further reduced to 828 and 693 on the fifth and seventh days, respectively. During the third day of culture, the amount of production of estradiol by FSH stimulation also decreased to about one-quarter of the first day's culture. Then, we created the FSHR expressing recombinant adenovirus. When it was introduced into the granulosa cells on the first day of culture, the expression of FSHR was enhanced up to 7,206 per cell and the amount of estradiol produced was remarkably increased up to 5,940 pg/ml on the third day. The co-culture of the immature oocytes with FSHR introduced granulosa cells is supposed to improve their developmental competence.