effects of fibroblast co-culture and activin a on in vitro growth of mouse preantral follicles

This study was conducted to evaluate fibroblast co-culture and Activin A on in vitro maturation and fertilization of mouse preantral follicles. The ovaries from 12-14-day-old mice were dissected, and 120-150 microm preantral follicles were cultured individually in Alpha-MEM as based medium for 12 days. Follicles were cultured in four conditions in a total number of 456 follicles: [i] base medium as control group [n = 113], [ii] base medium supplemented with 30 ng/ml Activin A [n = 115], [iii] base medium co-cultured with mouse embryonic fibroblast [n = 113], and [iv] base medium supplemented with 30 ng/ml Activin A and co-cultured with fibroblast [n = 115]. Rate of growth, survivability, antrum formation, ovulation, embryonic development and steroid production were evaluated. Analysis of Variance and Duncan test were applied for analyzing. Both co-culture and co-culture + Activin A groups showed significant difference [P<0.05] in growth [on days 4, 6, and 8 of culture period] and survival rates. However, there was no significant difference in antrum formation, ovulation rate, and embryonic development of ovulated oocytes. There were significant differences [P<0.05] in the estradiol secretion on days 8, 10, and 12 between co-culture + Activin A and the control group. Progesterone production also was significant [P<0.05] in co-culture + Activin A group on days 6, 8, 10, and 12 compared to control group. Fibroblast co-culture and Activin A promoted growth and survivability of preantral follicles. However, simultaneous use of them was more efficient

Effect of fibroblast co-culture on in vitro maturation and fertilization of mouse preantral follicles

The aim of this study was to evaluate fibroblast co-culture in vitro maturation and fertilization of prepubertal mouse preantral follicles. The ovaries of 12-14 day old mice were dissected and 120-150 micro m intact preantral follicles with one or two layers of granulose cells, and round oocytes were cultured individually in alpha-minimal essential medium [alpha-MEM] supplemented with 5% fetal bovine serum [FBS], 100 mIU/ml recombinant follicle stimulating hormone, 1% insulin, transferring, selenium mix, 100 micro g/ml penicillin and 50 micro 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. 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. Fibroblast co-culture increased survival rate and steroid production of preantral follicles by promoting granulose cell proliferation