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

Infertility in Mazandaran province - north of Iran: an etiological study

The prevalence and etiology of infertility are not similar in different parts of the world. There are only few reports of this topic in Iran. This study was conducted to determine the clinical patterns and major causes of infertility in Mazandaran province in north of Iran. The medical records of 3734 consecutive couples attending two infertility clinics in Mazandaran province, from 2003 to 2008, were reviewed. The couples had not had a viable birth after at least 1 year of unprotected intercourse and were fully investigated. Of the entire samples, 78.7% had primary infertility and 21.3% had secondary infertility. The mean duration of infertility in couples was 5.7 +/- 4 years. The etiology of infertility in couples revealed; male factor in 38.9%, female factor in 34.7%, combined factors in 14.6% and undetermined cause in 11.8%. In this study, delayed attendance of infertile couples to the infertility clinic was found. Therefore, there is a need to revise public health program on infertility to focus on the education and prevention of infertility and its risk factors

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

[Morphometrical study of mouse pre-implantation embryos: comparison of in vitro- and in vivo- produced embryos]

The development of pre-implantation mammalian embryos in vitro is compromised, compared with those grown in vivo. Selecting embryos with a high implantation potential is one of the most important challenges in the field of assisted reproductive technology. The aim of this study was to postulate morphometrical characteristics of good quality embryos, with comparisons between in vivo and in vitro produced mouse embryos. Embryos was obtained from NMRI female mice after super ovulation. In vivo developed 2-, 4- and 8-cell embryos; morulla and full blastocyst were isolated from mice on 18, 36, 52, 60, 72 and 96 hours after hCG administration respectively. Ham, s F10 medium was used for in vitro culture of embryos. External and internal diameter of embryos, zona thickness and number of cells in full blastocysts were evaluated and compared between in vivo and in vitro groups. External and internal diameter and zone thickness in oocyte and zygotes were 99.9microm, 75.4microm and 4.9microm respectively. These values did not change prior to the blastocyst stage in both in vivo and in vitro groups; but in full blastocyst stage, the diameter of embryos significantly increased and zone thickness decreased compared to prior stages in both groups [P<0.01]. The diameter of full blastocysts of in vivo group [116.5 microm] were significantly larger than those of in vitro group [104.3 microm, P<0.05]. Moreover, the full blastocysts of in vivo group had significantly more blastomeres [49], compared to in vitro group [43, P<0.05]. Additionally, cultured embryos reached full blastocyst at 110 hours after hCG administration, while in vivo condition the time frame was 96 hours. Based on the above results, embryo size and zona thickness cannot predict embryo quality prior to blastocyst stage, however, in this stage; larger embryos and those that have more blastomere may show greater viability