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  • Title: The optimized research of the in vitro culture of preantral follicles in mice.
    Author: Wang X, Wang L, Sun Y, Wu D, Li J, Zhu L, Jiang S, Pan X.
    Journal: J Clin Lab Anal; 2020 Nov; 34(11):e23498. PubMed ID: 33463764.
    Abstract:
    OBJECTIVE: Efficiency of preantral follicle culture in vitro is low and is dependent on species, development stage, and follicle-stimulating hormone (FSH) concentration. Here, we optimized the preantral follicle in vitro culture system in mice. METHODS: The primary follicles (PM follicles, 80-100 μm diameter ) and early secondary follicles (ES follicles, 110-130 μm diameter) isolated from 14-day female mice were cultured in mediums containing 10 mIU/mL or 100 mIU/mL r-FSH. The follicle growth and oocyte maturation were observed. Estradiol (E2) was detected by ELISA. FSH receptor (FSHR), Ki-67, 3β-HSD, CYP17, and CYP19 levels were detected by immunofluorescence and Western blot. RESULTS: The antrum formation and oocyte maturation rates of ES follicles were significantly higher than those of PM follicles (P < .05). They were also significantly higher in ES follicles with 100 mIU/mL r-FSH than with 10 mIU/mL r-FSH (P < .05). A higher FSHR level was found in ES follicles. Meanwhile, with 10 mIU/mL r-FSH, the ES follicles exhibited a pattern of flat growth, whereas a pattern of stereoscopic spatial growth was observed with 100 mIU/mL r-FSH. The 100 mIU/mL r-FSH stimulated granulosa cell proliferation more significantly than 10 mIU/mL r-FSH. Moreover, FSH significantly promoted ES follicle granulosa cell proliferation compared to PM follicular granulosa cells. The secretion of E2 and the expressions of 3β-HSD, CYP 17, and CYP 19 in ES follicles with 100 mIU/mL r-FSH were significantly higher than those with 10 mIU/mL r-FSH. CONCLUSIONS: The 100 mIU/mL r-FSH ideally promotes the development of ES follicles, whose growth pattern can more reasonably simulate the growth of follicles in vivo.
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