Author/Authors :
Heidari، Mahmoud نويسنده Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran , , Karimpour Malekshah، Abbasali نويسنده Department of Anatomy and Embryology, Molecular Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Science, Sari, , , Parivar، Kazem نويسنده , , Khanbabaei، Ramezan نويسنده Department of Biology, Qaemshahr Branch, Islamic Azad University, Qaemshahr, Iran , , Rafiei، Alireza نويسنده ,
Abstract :
Background: The aim of this study was to evaluate fibroblast co-culture on in vitro maturation and
fertilization of prepubertal mouse preantral follicles.
Materials and Methods: The ovaries of 12-14 day old mice were dissected and 120-150 ?m intact preantral follicles with one or two layers of granulosa cells, and round oocytes were cultured individually in ?-minimal essential medium (?-MEM) supplemented with 5% fetal bovine serum (FBS), 100 mIU/ml recombinant follicle stimulating hormone, 1% insulin, transferrin, selenium mix, 100 ?g/ml penicillin and 50 ?g/ml streptomycin as base medium for 12 days. A total number of
226 follicules were cultured under two conditions: i) base medium as control group (n=113); ii) base medium co-cultured with mouse embryonic fibroblast (MEF) (n=113). Follicular diameters, alone, in addition to other factors were analyzed by student’s t-test and chi-square test, respectively. Results: The co-culture group showed significant differences (p < 0.05) in growth rate (days 4, 6 and
8 of the culture period) and survival rate. However, there was no significant difference in antrum formation, ovulation rate and embryonic development of released oocytes. There were significant differences (p < 0.05) in the estradiol and progesterone secretion at all days between the co-culture and control groups.
Conclusion: Fibroblast co-culture increased survival rate and steroid production of preantral follicles by promoting granulosa cell proliferation.
