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Development of a serum‐free defined system employing growth factors for preantral follicle culture
Author(s) -
Park Young Hyun,
Gong Seung Pyo,
Kim Hwa Young,
Kim Gil Ah,
Choi Jun Hee,
Ahn Ji Yeon,
Lim Jeong Mook
Publication year - 2013
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.22204
Subject(s) - biology , follicle , blastocyst , oocyte , andrology , folliculogenesis , ovarian follicle , medicine , stem cell factor , chemically defined medium , endocrinology , fetal bovine serum , hair follicle , leukemia inhibitory factor , in vitro , embryo , stem cell , embryogenesis , ovary , immunology , microbiology and biotechnology , haematopoiesis , biochemistry , interleukin 6 , cytokine
SUMMARY This study was conducted to evaluate if mouse preantral follicles can yield developmentally competent oocytes following culture in serum‐free, defined medium. Donor follicles were obtained from 14‐day‐old B6CBAF1 mice, and cultured in α‐MEM‐Glutamax medium. The replacement of fetal bovine serum with knockout serum replacement (KSR) did not significantly reduce follicle growth or oocyte maturation in vitro, although it significantly reduced the development of oocytes after activation. Regardless of the replacement medium, follicle growth was not influenced by the addition of leukemia inhibitory factor (LIF). The addition of 100 ng/ml stem cell factor (SCF) to the KSR‐supplemented serum‐free medium significantly stimulated follicle development, which further improved blastocyst formation after oocyte activation. On Day 3 of culture, a significant increase in Bmp7 expression was detected in the SCF‐containing medium compared with the serum‐containing medium, whereas Gdf9 and Amh were increased in the serum‐containing medium. A significant increase in estradiol production was detected under serum‐free conditions, but minimal progesterone secretion was detected throughout the culture period. In conclusion, serum‐free media can be used to optimize ovarian follicle cultures, and the addition of SCF is beneficial for deriving developmentally competent oocytes through follicle culture. Mol. Reprod. Dev. 80: 725–733, 2013. © 2013 Wiley Periodicals, Inc .