Effects of a-MEM and TCM-199 culture media and epidermal growth factor on survival and growth of goat and sheep preantral follicles cultured in vitro

The aim of this study was to evaluate the effect of culture media (Alpha Minimum Essential Medium, α-MEM; and Tissue Culture Medium-199, TCM-199) in the absence or presence of Epidermal Growth Factor (EGF) on an in vitro culture of goat and sheep preantral follicles enclosed in ovarian tissue. The fragments of ovarian cortex from both species were immediately analyzed after collection (non-cultured control group) or cultured for 1 or 7 days in α-MEM+ or TCM-199+in the absence or presence of EGF (10 ng/ml). Before and after the culture, the fragments of ovarian cortex were analyzed by classical histology and fluorescence microscopy. After 1 day of culture, all treatments decreased the percentage of morphologically normal follicles when compared to non-cultured control in both species (P 0.05). Regarding follicle activation, an increase in the percentage of growing follicles was observed in all treatments after 7 days of culture when compared to the control group in both species. However, in sheep, after 7 days, only the treatments α-MEM+/EGF and TCM-199+showed larger(P < 0.05) oocytes than the control group. In conclusion, the TCM-199+ Preserved goat preantral follicle viability after in vitro culture. Furthermore, the media α-MEM+/EGF and TCM-199+ increased the oocyte diameter after 7 days of culture in sheep. Therefore, it isrecommended to use TCM-199+ In the culture of preantral follicles in both species.

Effects of a-MEM and TCM-199 culture media and epidermal growth factor on survival and growth of goat and sheep preantral follicles cultured in vitro

The aim of this study was to evaluate the effect of culture media (Alpha Minimum Essential Medium, α-MEM; and Tissue Culture Medium-199, TCM-199) in the absence or presence of Epidermal Growth Factor (EGF) on an in vitro culture of goat and sheep preantral follicles enclosed in ovarian tissue. The fragments of ovarian cortex from both species were immediately analyzed after collection (non-cultured control group) or cultured for 1 or 7 days in α-MEM+ or TCM-199+in the absence or presence of EGF (10 ng/ml). Before and after the culture, the fragments of ovarian cortex were analyzed by classical histology and fluorescence microscopy. After 1 day of culture, all treatments decreased the percentage of morphologically normal follicles when compared to non-cultured control in both species (P < 0.05). In fluorescence microscopy, viable sheep follicles were observed to decrease in all treatments after 7 days of culture when compared to non-cultured controls. However, in goats, the culture with TCM-199+maintained follicle viability after 7 days of culture, similar to fresh tissue (P > 0.05). Regarding follicle activation, an increase in the percentage of growing follicles was observed in all treatments after 7 days of culture when compared to the control group in both species. However, in sheep, after 7 days, only the treatments α-MEM+/EGF and TCM-199+showed larger(P < 0.05) oocytes than the control group. In conclusion, the TCM-199+ Preserved goat preantral follicle viability after in vitro culture. Furthermore, the media α-MEM+/EGF and TCM-199+ increased the oocyte diameter after 7 days of culture in sheep. Therefore, it isrecommended to use TCM-199+ In the culture of preantral follicles in both species.(AU)

Steady-state levels of vasoactive intestinal peptide (VIP) mRNA in goat ovaries and the effect of VIP on the in vitro development of isolated preantral follicles

The aims of this study were to verify the steady-state level of vasoactive intestinal peptide (VIP) mRNA in goat follicles at various developmental stages and to investigate the influence of VIP on the survival, antrum formation and growth of secondary follicles cultured for 6 days. Primordial, primary and secondary goat follicles and small and large antral follicles were obtained to quantify VIP mRNA by real-time reverse transcription with the polymerase chain reaction. The influence of VIP and the presence or absence of follicle-stimulating hormone (FSH) on the development of secondary follicles and on mRNA expression for VIP and FSH receptor (FSHR) were determined after 6 days of culture. Survival, antrum formation and follicular diameter were evaluated every other day of culture. The levels of VIP mRNA in primary and secondary follicles were significantly higher than in primordial follicles. Cumulus–oocyte complexes (COCs) from both small and large antral follicles had significantly higher levels ofVIP mRNA than their respective granulosa/theca cells. During culture, the addition of VIP and/or FSH had o effect on follicular development. However, the presence of FSH and/or VIP in the culture medium significantly reduced VIP mRNA levels, but did not alter FSHR mRNA levels. In conclusion, VIP mRNA was detected in all goat follicular categories and cellular types, VIP and/or FSH did not affect the development of secondary follicles and reduce the expression of VIP mRNA levels.(AU)

Steady-state levels of vasoactive intestinal peptide (VIP) mRNA in goat ovaries and the effect of VIP on the in vitro development of isolated preantral follicles

The aims of this study were to verify the steady-state level of vasoactive intestinal peptide (VIP) mRNA in goat follicles at various developmental stages and to investigate the influence of VIP on the survival, antrum formation and growth of secondary follicles cultured for 6 days. Primordial, primary and secondary goat follicles and small and large antral follicles were obtained to quantify VIP mRNA by real-time reverse transcription with the polymerase chain reaction. The influence of VIP and the presence or absence of follicle-stimulating hormone (FSH) on the development of secondary follicles and on mRNA expression for VIP and FSH receptor (FSHR) were determined after 6 days of culture. Survival, antrum formation and follicular diameter were evaluated every other day of culture. The levels of VIP mRNA in primary and secondary follicles were significantly higher than in primordial follicles. Cumulus–oocyte complexes (COCs) from both small and large antral follicles had significantly higher levels ofVIP mRNA than their respective granulosa/theca cells. During culture, the addition of VIP and/or FSH had o effect on follicular development. However, the presence of FSH and/or VIP in the culture medium significantly reduced VIP mRNA levels, but did not alter FSHR mRNA levels. In conclusion, VIP mRNA was detected in all goat follicular categories and cellular types, VIP and/or FSH did not affect the development of secondary follicles and reduce the expression of VIP mRNA levels.