Influence of the ovarian fragmentation before storage at 4o C on the apoptosis rates and in vitro development of ovine preantral follicles

The aim of this study was to evaluate the effect of ovarian tissue fragmentation before preservation at 4o C in MEM on the morphology, apoptosis, and growth of ovine preantral follicles. After collection, the ovaries were divided into two halves, being one divided into two fragments (1/4 of the ovary). One fragment was subdivided into two fragments (1/8), being one fixed for histology (fresh control). The remaining whole ovary, 1/2, 1/4 and 1/8 of the ovary were preserved in MEM at 4°C for 6, 12 or 24 h. The tissue was further destined to histology. In vitro culture and TUNEL technique were performed in treatments that showed the best results of follicular survival after preservation. Storage of 1/8 of the ovary increased the normal follicles compared with half or whole ovary. After preservation in 1/8 of the ovary and culture for 7 days, the percentage of apoptotic cells was similar to the fresh control and non-cultured fragments. The percentage of growing follicles increased after preservation of 1/8 of the ovary compared with 1/4. In conclusion, ovine preantral follicles can be preserved in fragments of 1/8 of the ovary in MEM at 4°C for 24 h, without affecting apoptosis and their ability to grow in vitro.

Influence of the ovarian fragmentation before storage at 4o C on the apoptosis rates and in vitro development of ovine preantral follicles

The aim of this study was to evaluate the effect of ovarian tissue fragmentation before preservation at 4o C in MEM on the morphology, apoptosis, and growth of ovine preantral follicles. After collection, the ovaries were divided into two halves, being one divided into two fragments (1/4 of the ovary). One fragment was subdivided into two fragments (1/8), being one fixed for histology (fresh control). The remaining whole ovary, 1/2, 1/4 and 1/8 of the ovary were preserved in MEM at 4°C for 6, 12 or 24 h. The tissue was further destined to histology. In vitro culture and TUNEL technique were performed in treatments that showed the best results of follicular survival after preservation. Storage of 1/8 of the ovary increased the normal follicles compared with half or whole ovary. After preservation in 1/8 of the ovary and culture for 7 days, the percentage of apoptotic cells was similar to the fresh control and non-cultured fragments. The percentage of growing follicles increased after preservation of 1/8 of the ovary compared with 1/4. In conclusion, ovine preantral follicles can be preserved in fragments of 1/8 of the ovary in MEM at 4°C for 24 h, without affecting apoptosis and their ability to grow in vitro.(AU)

Effect of ovarian tissue transportation in Amburana cearensis extract on the morphology and apoptosis of goat preantral follicles

The aim of the present study was to evaluate the effect of Amburana cearensis extract during caprine ovarian tissue transportation on the survival of preantral follicles in vitro. HPLC was used to determine the fingerprint chromatogram of the ethanolic extract. Five goat ovarian pairs were divided into fragments. One fragment was fixed for histology and TUNEL Analysis (fresh control). The other fragments were placed in MEM or A. cearensis extract (0.1; 0.2 or 0.4 mg/ml) and stored at 4oC for 6, 12 or 24 h. Preserved Fragments were also fixed for histology and TUNEL analysis. The presence of phenolic compounds (protocatechuic acid, epicatechin, p-coumaric acid, gallic acid and kaempferol) in the extract was confirmed using HPLC. Thepercentage of normal follicles preserved in 0.2 mg/ml A. cearensis for 6 h was similar to that observed in the fresh control. Moreover, the percentage of normal follicles was higher after preservation in 0.2 mg/ml A. cearensisFor 6 h than the other A. cearensis treatments and similar to that found in MEM. There were no differences in the percentage of apoptotic cells between fresh control and those preserved for 6 h in MEM or 0.2 mg/ml A. cearensis. In conclusion, both 0.2 mg/ml A. cearensis or MEM can be used for the preservation of goat preantral follicles for up to 6 h. The use of A. cearenses is recommended due to the higher cost of MEM.

Effect of ovarian tissue transportation in Amburana cearensis extract on the morphology and apoptosis of goat preantral follicles

The aim of the present study was to evaluate the effect of Amburana cearensis extract during caprine ovarian tissue transportation on the survival of preantral follicles in vitro. HPLC was used to determine the fingerprint chromatogram of the ethanolic extract. Five goat ovarian pairs were divided into fragments. One fragment was fixed for histology and TUNEL Analysis (fresh control). The other fragments were placed in MEM or A. cearensis extract (0.1; 0.2 or 0.4 mg/ml) and stored at 4oC for 6, 12 or 24 h. Preserved Fragments were also fixed for histology and TUNEL analysis. The presence of phenolic compounds (protocatechuic acid, epicatechin, p-coumaric acid, gallic acid and kaempferol) in the extract was confirmed using HPLC. Thepercentage of normal follicles preserved in 0.2 mg/ml A. cearensis for 6 h was similar to that observed in the fresh control. Moreover, the percentage of normal follicles was higher after preservation in 0.2 mg/ml A. cearensisFor 6 h than the other A. cearensis treatments and similar to that found in MEM. There were no differences in the percentage of apoptotic cells between fresh control and those preserved for 6 h in MEM or 0.2 mg/ml A. cearensis. In conclusion, both 0.2 mg/ml A. cearensis or MEM can be used for the preservation of goat preantral follicles for up to 6 h. The use of A. cearenses is recommended due to the higher cost of MEM.(AU)

Transforming growth factor-β (TGF-β) maintains follicular ultrastructure and stimulates preantral follicle growth in caprine ovarian tissue cultured in vitro / Fator de crescimento transformante-β (TGF-β) mantém a ultraestrutura folicular e estimula o crescimento de folículos pré-antrais caprinos inclusos em tecido ovariano cultivado in vitro

The objectives of this study were to investigate whether TGF-β affect the survival, activation and further growth of goat primordial follicles enclosed in ovarian cortex after in vitro culture. Goat ovaries were collected from an abattoir and pieces of ovarian tissues were cultured for one or seven days in a supplemented alpha Minimum Essential Medium, alone or containing TGF-β (1, 5, 10 or 50ng/mL). Ovarian tissues from the fresh control as well as those cultured were processed for histological and ultrastructural studies. The results showed that when compared with fresh control, there was decrease in the percentages of histologically normal follicles in all treatments only after seven days culture. TGF-β did not affect the activation of preantral follicles regardless of its concentration, however, larger follicles diameter (P<0.05) was observed using 10ng/mL TGF-β than in the fresh control and other treatments. Moreover, this concentration maintained the normal ultrastructure after seven days of culture. In conclusion, TGF-β showed additional effect on the follicle growth and the maintenance of ultrastructural integrity of goat preantral follicles enclosed in ovarian tissue when used at 10ng/mL during seven days of culture.(AU)

O objetivo desse estudo foi investigar se o TGF-β afeta a sobrevivência, ativação e crescimento de folículos primordiais caprinos inclusos no córtex ovariano após o cultivo in vitro. Ovários de cabras foram coletados em abatedouro e fragmentos de tecido ovariano foram cultivados por um e sete dias em meio essencial mínimo alfa (α-MEM+) sozinho ou suplementado com TGF-β (1, 5, 10 ou 50ng/mL). Fragmentos ovarianos não cultivados e cultivados foram processados para análise histológica e ultraestrutural. Os resultados mostraram que, comparado ao controle fresco, houve diminuição no percentual de folículos morfologicamente normais em todos os tratamentos somente após sete dias de cultivo. O TGF-β não afetou a ativação folicular independente da concentração testada, contudo, o diâmetro folicular foi superior (P<0.05) no tratamento com 10ng/mL de TGF-β quando comparado ao controle fresco e aos demais tratamentos. Além disso, essa mesma concentração manteve a ultraestrutura normal dos folículos após sete dias de cultivo. Em conclusão, o TGF-β apresentou efeito adicional no crescimento folicular e na manutenção da integridade ultraestrutural de folículos pré-antrais caprinos inclusos no tecido ovariano quando utilizado na concentração de 10ng/mL durante sete dias de cultivo.(AU)