Effect of base media, FSH and anti-Müllerian hormone (AMH) alone or in combination on the growth of pig preantral follicles in vitro





Culture; Esteroids; Folliculogenesis; Porcine.


To compare the efficiency of North Carolina State University medium 23 (NCSU23) and Alpha Minimum Essential Medium (α-MEM) as a base medium, and to evaluate the effects of Anti-Müllerian Hormone (AMH) alone or in combination with Follicle Stimulating Hormone (FSH) on the in vitro development and steroid production of isolated porcine preantral follicles. Porcine secondary follicles were cultured in NCSU23 or α-MEM media for 4 days. Once α-MEM was determined to be the optimal culture medium, secondary follicles were cultured in α-MEM alone or supplemented with FSH (1.5 ng/mL), AMH (50 ng/mL) or the combination of the two hormones. Follicle development was evaluated by measuring follicular growth, morphology and hormone production. There was no difference between the media NCSU23 and α-MEM in terms of follicle survival and growth (P > 0.05). However, at day 2, the antrum formation rate tended to be (P < 0.074) higher in α-MEM than NCSU23. At day 4 of culture, the estradiol and progesterone secretion were higher in α-MEM than NCSU23 (P < 0.01), while the opposite was observed for testosterone (P < 0.01). The addition of AMH and/or FSH did not affect follicular survival and growth. Nevertheless, the secretion of estradiol and progesterone induced by FSH was reduced with AMH (P < 0.01). α-MEM is a more effective base medium than NCSU23 for the in vitro follicular development of pig preantral follicles and AMH reduces the steroid production induced by FSH.


Cao, Z., Gao, D., Tong, X., Xu, T., Zhang, D., Wang, Y., Liu, Y., Li, Y., Zhang, Y., & Pu, Y. (2019). Melatonin improves developmental competence of oocyte-granulosa cell complexes from porcine preantral follicles. Theriogenology, 133, 149-158.

Carlsson, I. B., Scott, J. E., Visser, J. A., Ritvos, O., Themmen, A. P., & Hovatta, O. (2006). Anti-Müllerian hormone inhibits initiation of growth of human primordial ovarian follicles in vitro. Human Reproduction, 21, 2223-2227.

Chang, H. M., Klausen, C., & Leung, P. C. (2013). Anti-müllerian hormone inhibits follicle-stimulating hormone-induced adenylyl cyclase activation, aromatase expression, and estradiol production in human granulosa-lutein cells. Fertility and Sterility, 100, 585-592.

Dewailly, D., Robin, G., Peigne, M., Decanter, C., Pigny, P., & Catteau-Jonard, S. (2016). Interactions between androgens, FSH, anti-Müllerian hormone and estradiol during folliculogenesis in the human normal and polycystic ovary. Human Reproduction Update, 22, 6, 709-724.

Di Clemente, N., Wilson, C., Faure, E., Boussin, L., Carmillo, P., Tizard, R., Picard, J. Y., Vigier, B., Josso, N., & Cate, R. (1994). Cloning, expression, and alternative splicing of the receptor for anti-Müllerian hormone. Molecular Endocrinology, 1006-1020.

Durlinger, A. L., Gruijters, M. J., Kramer, P., Karels, B., Kumar, T. R., Matzuk, M. M., Rose, U. M., Jong, F. H., Uielenbroek, J. T., Grootegoed, J. A., & Themmen, A. P. (2001). Anti-Müllerian hormone attenuates the effects of FSH on follicle development in the mouse ovary. Endocrinology, 142, 4891-4899.

Durlinger, A. L. L., Gruijters, M. J. G., Kramer, P., Karels, B., Ingraham, H. A., Nachtigal, M. W., Uilenbroek, J. T. J., & Grootegoed, J. A. (2002). Anti-Müllerian hormone inhibits initiation of primordial follicle growth in the mouse ovary. Endocrinology, 143, 1076-1084.

Figueiredo, J. R., Rodrigues, A. P. R., Silva, J. R. V., & Santos, R. R. (2011). Cryopreservation and in vitro culture of caprine preantral follicles. Reproduction, Fertility and Development, 23, 1, 40-47.

Kano, M., Sosulski, A. E., Zhang, L., Saatcioglu, H. D., Wang, D., Nagykery, N., Sabatini, M. E., Gao, G., & Donahoe, P. K. (2017). AMH/MIS as a contraceptive that protects the ovarian reserve during chemotherapy. Proceedings of the National Academy of Sciences, 114, 9, E1688-E1697.

Lima, L. F., Rubess, M., Rocha, R., Winters, R., Milner, D. J., Campello, C. C., Figueiredo, J. R., & Wheeler, M. B. (2017). High diluted and dynamised follicle stimulating hormone modulates steroid production in isolated porcine preantral follicles cultured in vitro. Homeopathy, 106, 2, 87-92.

Linardi, A., Damiani, D., & Longui, C. A. (2017). The use of aromatase inhibitors in boys with short stature: what to know before prescribing? Arch Endocrinology and Metabolism, 61, 4, 391-397.

Mao, J., Wu, G., Smith, M.F., McCauley, T. C., Cantley, T. C., Prather, R. S., Didion, B. A., & 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. Biology of Reproduction, 67, 1197-1203.

Mao, J., Smith, M. F., Rucker, E. B., Wu, G. M., McCauley, T. C., Cantley, T. C., Prather, R. S, Didion, B. A., & Day, B. N. (2004). Effect of epidermal growth factor and insulin-like growth factor I on porcine preantral follicular growth, antrum formation, and stimulation of granulosa cell proliferation and suppression of apoptosis in vitro. Journal of Animal Science, 82, 7, 1967-1975.

Marino, P. C., Bizarro-Silva, C., Búfalo, I., Rosa, C. O., Gonçalves, G. R., Lisboa, L. A., & Seneda, M. M. (2019). Growth and Differentiation Factor-9 Supplementation Affects Viability and Morphology of Preantral Follicles in Equine Ovarian Fragments During Short-term In vitro Culture. Brazilian Archives of Biology and Technology, 62.

Nilsson, E. E., Schindler, R., Savenkova, M. I., & Skinner, M. K. (2011). Inhibitory actions of Anti-Müllerian Hormone (AMH) on ovarian primordial follicle assembly. PloS One, 5, e20087.

O’Brien, M. J., Pendola, J. K., & Eppig, J. J. (2003). A revised protocol for in vitro development of mouse oocytes from primordial follicles dramatically improves their developmental competence. Biology of Reproduction, 68, 5, 1682-1686.

Park, J. H., Maclaughlin, D. T., & Teixeira, J. M. (2011). The rate of in vitro maturation of primary follicles from adult mice and the quality of oocytes is improved in the absence of anti-mullerian hormone. Reproductive Sciences, 18, 334-341.

Pankhurst, M. W. (2017). A putative role for anti-Müllerian hormone (AMH) in optimising ovarian reserve expenditure. Journal Endocrinology, 233, 1, R1-R13.

Pankhurst, M. W., Kelley, R. L., Sanders, R.L., Woodcock, S. R., Oorschot, D. E., & Batchelor, N. J. (2018). Anti-Müllerian hormone overexpression restricts preantral ovarian follicle survival. Journal Endocrinology, 237, 2, 153-163.

Pellatt, L., Rice, S., Dilaver, N., Heshri, A., Galea, R., Brincat, M., Brown, K., Simpson, E. R., & Mason, H. D. (2011). Anti-Müllerian hormone reduces follicle sensitivity to follicle-stimulating hormone in human granulosa cells. Fertility and Sterility, 96, 1246-1251.

Pyoos, G. M., Maqhashu, A. M., Scholtz, M. M., & Nedambale, T. L. (2018). The comparison of three media on the in vitro maturation rate of pig oocytes. South African Journal of Animal Science, 48, 1027.

Rocha, R. M. P., Lima, L. F., Brito, I. R., Silva, G. M., Correia, H. H., Sá, N. A. R., Ferreira, A. C. A., Sales, A. D., Lobo, C. H., Campello, C. C., Smitz, J. S., Wheeler, M. B., & Figueiredo, J. R. (2016). Anti-Müllerian hormone reduces growth rate without altering follicular survival in isolated caprine preantral follicles cultured in vitro. Reproduction, Fertility and Development, 29, 1144-1154.

Rubessa, M., Rocha, R., Lima, L., Winters, R., Figueiredo, J., & Wheeler, M. (2016). 136 Comparison of NCSU-23 and Alpha-Minimal Essential Media in the Development of Isolated Porcine Preantral Follicles In vitro. Reproduction, Fertility and Development, 28, 1, 198.

Sacchi, S., D'Ippolito, G., Sena, P., Marsella, T., Tagliasacchi, D., Maggi, E., Argento, C., Tirelli, A., Giulini, S., La, & Marca. A. (2016). The anti-Müllerian hormone (AMH) acts as a gatekeeper of ovarian steroidogenesis inhibiting the granulosa cell response to both FSH and LH. Journal of Assisted Reproduction and Genetics, 33, 1, 95-100.

Schmidt, K. L., Kryger-Baggesen, N., Byskov, A. G., & Andersen, C. Y. (2005). Anti-Müllerian hormone initiates growth of human primordial follicles in vitro. Molecular and Cellular Endocrinology, 234, 87-93.

Sonigo, C., Beau, I., Grynberg, M., & Binart, N. (2019). AMH prevents primordial ovarian follicle loss and fertility alteration in cyclophosphamide-treated mice. The FASEB Journal, 33, 1, 1278-1287.

Tasaki, H., Iwata, H., Sato, D., Monji, Y., & Kuwayama, T. (2013). Estradiol has a major role in antrum formation of porcine preantral follicles cultured in vitro. Theriogenology, 79, 809-814.

Valdez, K. E., & Turzillo, A. M. (2005). Regulation of nuclear factor-kappaB (NF-kappaB) activity and apoptosis by estradiol in bovine granulosa cells. Molecular and Cellular Endocrinology, 243, 66-73.

Wu, J., Emery, B. R., & Carrell, D. T. (2001). In vitro growth, maturation, fertilization, and embryonic development of oocytes from porcine preantral follicles. Biology of Reproduction, 64, 1, 375-381.

Wu, J., Xu, B., & Wang, W. (2007). Effects of luteinizing hormone and follicle stimulating hormone on the developmental competence of porcine preantral follicle oocytes grown in vitro. Journal of Assisted Reproduction and Genetics, 24, 419-424.




How to Cite

ROCHA, R. M. P. .; RUBESSA, M. .; LIMA, L. F. de .; SILVA, A. F. B. da .; WINTERS, R.; POLKOFF, K. .; MILNER, D. .; CAMPELLO, C. C. .; FIGUEIREDO, J. R. de; WHEELER, M. . Effect of base media, FSH and anti-Müllerian hormone (AMH) alone or in combination on the growth of pig preantral follicles in vitro. Research, Society and Development, [S. l.], v. 10, n. 15, p. e53101522488, 2021. DOI: 10.33448/rsd-v10i15.22488. Disponível em: https://rsdjournal.org/index.php/rsd/article/view/22488. Acesso em: 5 dec. 2023.



Agrarian and Biological Sciences