Role of Follicle Stimulating Hormone in the Survival, Activation and Further Growth of in vitro Cultured Sheep Primordial Follicles

Document Type : Research Article


1 Department of Animal Science, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Reproduction and Development, Reproductive Biomedicine Center, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran


The aim of the present study was to investigate the effect of follicle stimulating hormone (FSH) on survival, activation and growth of ovine primordial follicles using histological studies. Pieces of ovine ovarian cortex were obtained and cultured for 6 days in Minimum Essential Medium supplemented with or without FSH (50 ng/mL). These fragments were then process to be used for histology compared to each other and non-cultured group. The results showed that the lowest percentage of normal follicles in control medium was observed after 6 days of culture. A higher percentage of growing follicles was observed in presence vs. absence of FSH. Moreover, an increase in diameter of both oocyte and follicle was observed in those fragments that have been cultured in presence of FSH. This study demonstrated that FSH not only maintains the morphological integrity of ovine preantral follicles cultured for 6 days in vitro, but also stimulates the activation of primordial follicles and the growth of activated follicles.


Adriaens I., Cortvrindt R. and Smitz J. (2004). Differential FSH exposure in preantral follicle culture has marked effects on folliculogenesis and oocyte developmental competence. Hum. Reprod. 19, 398-408.
Andrade E.R., Maddox-Hyttel P., Da Cruz Landim-Alvarenga F., Silva J.R.V., Alfieri A.A., Seneda M.M., Figueiredo J.R. and Toniolli R. (2011). Ultrastructure of sheep primordial follicles cultured in the presence of indol acetic Acid, EGF, and FSH. Vet. Med. Int. 7(2), 80-89.
Andrade E.R., Seneda M.M., Alfieri A.A., Oliveira J.A., Bracarense A.P.F.R.L., Figueiredo J.R. and Toniolli R. (2005). Interactions of indole acetic acid with EGF and FSH in the culture of ovine preantral follicles. Theriogenology. 64, 1104-1113.
Chun S.Y., Billig H., Tilly J.L., Furuta I., Tsafriri A. and Hsueh A.J. (1994). Gonadotropin suppression of apoptosis in cultured preovulatory follicles: mediatory role of endogenous insulin-like growth factor I. Endocrinology. 135, 1845-1853.
Cortvrindt R., Smitz J. and Van Steirteghem A.C. (1997). Assessment of the need for follicle stimulating hormone in early preantral mouse follicle culture in vitro. Hum. Reprod. 12, 759-768.
Dierich A., Sairam M.R., Monaco L., Fimia G.M., Gansmuller A., LeMeur M. and Sassone-Corsi P. (1998). Impairing follicle-stimulating hormone (FSH) signaling in vivo: targeted disruption of the FSH receptor leads to aberrant gametogenesis and hormonal imbalance. Proc. Natl. Acad. Sci. USA. 95, 13612-13617.
Durlinger A.L., Gruijters M.J., Kramer P., Karels B., Ingraham H.A., Nachtigal M.W., Uilenbroek J.T., Grootegoed J.A. and Themmen A.P. (2002). Anti-mullerian hormone inhibits initiation of primordial follicle growth in the mouse ovary. Endocrinology. 143, 1076-1084.
Hsueh A.J., Billig H. and Tsafriri A. (1994). Ovarian follicle atresia: a hormonally controlled apoptotic process. Endocrinol. Rev. 15, 707-724.
Itoh T., Kacchi M., Abe H., Sendai Y. and Hoshi H. (2002). Growth, antrum formation, and estradiol production of bovine preantral follicles cultured in a serum-free medium. Biol. Reprod. 67, 1099-1105.
Lima-Verde I.B., Rossetto R., Matos M.H.T., Celestino J.J.H., Bruno J.B., Silva C.M.G., Faustino L.R., Mororó M.B.S., Araújo V.R., Campello C.C. and Figueiredo J.R. (2010). Androstenedione and follicle stimulating hormone involvement in the viability and development of goat preantral follicles in vitro. Anim. Reprod. 7(2), 80-89.
Magalhães D.M., Araújo V.R., Lima-Verde I.B., Matos M.H.T., Silva R.C., Lucci C.M., Báo S.N., Campello C.C. and Figueiredo J.R. (2009). Impact of pituitary FSH purification on in vitro early folliculogenesis in goats. Biocell. 33, 91-97.
Mao J., Wu G., Smith M.F., McCauley T.C., Cantley T.C., Prather R.S., Didion B.A. and Day B.N. (2002). Effects of culture medium, serum type, and various concentrations of follicle-stimulating hormone on porcine preantral follicular development and antrum formation in vitro. Biol. Reprod. 67, 1197-1203.
Matos M.H., Lima-Verde I.B., Bruno J.B., Lopes C.A., Martins F.S., Santos K.D., Rocha R.M., Silva J.R., Báo S.N. and Figueiredo J.R. (2007a). Follicle stimulating hormone and fibroblast growth factor-2 interact and promote goat primordial follicle development in vitro. Reprod. Fertil. Dev. 19(5), 677-684.
Matos M.H., Lima-Verde I.B., Luque M.C., Maia J.E., Silva J.R., Celestino J.J., Martins F.S., Báo S.N., Lucci C.M. and Figueiredo J.R. (2007b). Essential role of follicle stimulating hormone in the maintenance of caprine preantral follicle viability in vitro. Zygote. 15(2), 173-182.
McGee E., Spears N., Minami S., Hsu S.Y., Chun S.Y., Billig H. and Hsueh A.J.W. (1997). Preantral ovarian follicles in serum-free culture: suppression of apoptosis after activation of the cyclic guanosine 3’-5’-monophosphate pathway and stimulation of growth and differentiation by follicle-stimulating hormone. Endocrinology. 138, 2417-2424.
Méduri G., Charnaux N., Driancourt M.A., Combettes L., Granet P., Vannier B., Loosfelt H. and Milgrom E. (2002). Follicle-stimulating hormone receptors in oocytes? J. Clin. Endocrinol. Metab. 87, 2266-2276.
O’Brien M.J., Pendola J.K. and Eppig J.J. (2003). A revised protocol for in vitro development of mouse oocytes from primordial follicles dramatically improves their developmental competence. Biol. Reprod. 68, 1682-1686.
Oktay K., Briggs D. and Gosden R.G. (1997). Ontogeny of follicle-stimulating hormone receptor gene expression in solated human ovarian follicles. J. Clin. Endocrinol. Metab. 82, 3748-3751.
Oktay K., Newton H., Mullan J. and Gosden R.G. (1998). Development of human primordial follicles to antral stages in SCID/hpg mice stimulated with follicle stimulating hormone. Hum. Reprod. 13, 1133-1138.
O'Shaughnessy P.J., Dudley K. and Rajapaksha W.R. (1996). Expression of follicle stimulating hormone-receptor mRNA during gonadal development. Mol. Cell. Endocrinol. 125, 169-175.
Peng X., Yang M., Wang L., Tong C. and Guo Z. (2010). In vitro culture of sheep lamb ovarian cortical tissue in a sequential culture medium. J. Assist. Reprod. Genet. 101, 30-41.
Picton H.M., Harris S.E., Muruvi W. and Chambers E.L. (2008). The in vitro growth and maturation of follicles. Reproduction. 136, 703-715.
Saha S., Shimizu M., Geshi M. and Izaike Y. (2000). In vitro culture of bovine preantral follicles. Anim. Reprod. Sci. 63, 27-39.
Silva J.R.V., Van den Hurk R., Matos M.H.T., Santos R.R., Pessoa C., Moraed M.O. and Figueiredo J.R. (2004). Influences of FSH and EGF on primordial follicles during in vitro culture of caprine ovarian cortical tissue. Theriogenology. 61, 1691-1704.
Thomas F.H., Ethier J.F., Shimasaki S. and Vanderhyden B.C. (2005). Follicle-stimulating hormone regulates oocyte growth by modulation of expression of oocyte and granulosa cell factors. Endocrinology. 146, 941-949.
Touraine P., Beau I., Gougeon A., Meduri G., Desroches A., Pichard C., Detoeuf M., Paniel B., Prieur M., Zorn J.R., Milgrom E., Kutten F. and Misrahi M. (1999). New natural inactivating mutations of the follicle-stimulating hormone receptor: correlations between receptor function and phenotype. Mol. Endocrinol. 13, 1844-1854.
Van den Hurk R. and Zhao J. (2005). Formation of mammalian oocytes and their growth, differentiation and maturation within ovarian follicles. Theriogenology. 63, 1717-1751.
Yu-bin L., Can-quan Z., Guo-fen Y., Qiong W. and Yu D. (2007). Modified vitrification method for cryopreservation of human ovarian tissues. Chin. Med. J. 120(2), 110-114.
Zhou H. and Zhang Y. (2005). Regulation of in vitro growth of preantral follicles by growth factors in goats. Domest. Anim. Endocrinol. 28, 235-242.