Effects ofin vitroculture of mouse fetal gonads on subsequent ovarian developmentin vivoand oocyte maturationin vitro

Abstract

Under organ culture, female fetal gonads in mice cannot develop beyond the preantral follicle stage unless the follicles are individually isolated and cultured again. In this study, we investigated the effect of in vitro culture of female fetal gonads before transplantation on subsequent in vivo development. The gonads derived from female fetuses 12.5 days postcoitum were organ-cultured for 0, 7 and 14 days, and then were grafted underneath the kidney capsules of severe combined immunodeficient mice and recovered at 21, 14 and 7 days post-transplantation, respectively. The histological analysis of the grafts showed that the in vitro culture of the fetal gonads restricted follicular development to the antral follicle stage post-transplantation. In the grafts cultured for 14 days, particularly, no antral follicle was observed. However, the oocytes in these follicles had grown to around 65 microm in diameter and had competence to resume meiosis in vitro. When the fetal gonads were grafted after culture for 7 and 14 days, 13.0% and 6.8% of the oocytes progressed to the metaphase II stage, respectively. These data showed significant differences (P < 0.05) in comparison with the control group (25.3%). Our results indicate that the in vitro culture of female fetal gonads before transplantation affects the subsequent in vivo development of both follicular cells and oocytes, and in vitro oocyte maturation. However, this effect seems to be more severe in terms of follicular development when compared with oocyte growth and maturation.

Follicle growth and oocyte development after ovary transplantation into back muscle of immune-intact adult castrated male mice

“…Follicle and oocyte development after grafting used for ovarian transplantation (Candy et al 2000, Hosoe et al 2008, Schubert et al 2008, Soleimani et al 2008, Abir et al 2009, Motohashi et al 2009). For the immune-intact animals as recipients, immunosuppressant treatment is the first and most effective choice.…”

“…The recovered oocytes were rinsed, and then matured in vitro in a-MEM medium supplemented with 10 IU/ml eCG, 10 IU/ml hCG, 10 ng/ml epidermal growth factor, and 5% FBS (Motohashi et al 2009) at 37 8C in 5% CO 2 in air. Sixteen hours later, the PB1 extrusion was examined.…”

“…Immunotolerance was also not induced between mouse strains (Gosden 2007(Gosden , 2008. So far, in order to facilitate these experiments, most studies have used immunodeficient or inbred mice as recipients to reduce/eliminate rejection after ovarian transplantation (Candy et al 2000, Hosoe et al 2008, Schubert et al 2008, Soleimani et al 2008, Abir et al 2009, Motohashi et al 2009). In these cases, athymic nude mice or severe combined immunodeficient mice (who exhibition absence of mature B and T cells) were used to prevent immune rejection.…”

Recovery of Functional Oocytes from Cultured Premeiotic Germ Cells After Kidney Capsule Transplantation

“…The adult ectopic in vivo environment, such as the kidney capsule, supports the maturation of oocytes from purified fetal germ cells through aggregation and transplantation procedures [45], and living offspring had obtained from 12.5 dpc fetal mouse ovaries after ectopic transplantation [46]. Notably, recipient mice seem able to support the full-term development of female germ cells, despite sex differences in the hormonal environment of the recipients [46,47]. Finally, both embryonic and adult stem cell-derived germ cells obtained in vitro appear able to form developing follicles after aggregation with ovarian somatic cells and transplanted beneath the kidney capsule of recipients [28,48].…”

“…Grafts were collected from the recipient mice at the day described. Five IU pregnant mare serum gonadotropin (PMSG) was intraperitoneally injected to the recipient mice 48 h before collection [47].…”