2011 (SBTE) 25th Annual Meeting Proceedings - International ...

Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011.
A041 FOLLICULOGENESIS, OOGENESIS AND SUPEROVULATION
ADDITION OF INSULIN TO THE IN VITRO CULTURE MEDIUM PROMO
OMOTES SURVIV
VIVAL AND DEVEL
VELOPMENT OF FOLLICLES
PREANTRAL AL GOATS
Roberta Nogueira Chaves, , Anelise Maria Costa Vasconcelos Alves, Luciana Rocha Faustino, Claudio Afonso Pinho Lopes, Kenio Patricio Lima Oliveira & José
Ricardo de Figueiredo
LABORATÓRIO DE MANIPULAÇÃO DE OÓCITOS E FOLÍCULOS PRÉ-ANTRAIS (LAMOFOPA), UECE, FORTALEZA, CE, BRAZIL.
Insulin is usually added to the ovarian tissue culture as a factor that stimulates cell survival and mitosis, but no studies to
determine the best concentration to be used. Thus, the aim of this study was to elucidate the effects of the addition of different insulin
concentrations to the culture medium on the survival and development of goat preantral follicles after 7 days of culture. Ovaries (n=14) from
adult, non-pregnant, mixed-breed goats (1-3 years of age) were collected from a local slaughterhouse. In the laboratory, the cortex from each
ovarian pair was sliced and the fragments were fixed (fresh control) or cultured for 1 or 7 days in the absence or presence of insulin (0, 5, 10
ng/mL and 5 or 10 µg/mL). Each treatment was repeated seven times. Non-cultured and cultured tissues were processed for classical histology,
transmission electron microscopy and viability test using fluorescent probes (calcein-AM and ethidium homodimer-1). The data were
subjected to analysis of variance and compared using the Dunnett’s, SNK and ÷2 tests (P < 0.05). After 7 days of culture in medium
supplemented with 10 ng/mL insulin, 82% of the follicles present are morphologically normal. This percentage was significantly higher than
those observed in control medium (71%) or supplemented with 5 (65%) and 10 µg/mL (66%), but did not differ from follicles cultured with
5 ng/mL (74%) of insulin. The ultrastructural analysis and the viability test confirmed the integrity of follicles cultured for 7 days in medium
containing 10 ng/mL of insulin. At the end of culture, all treatments significantly increased the percentage of developing follicles (0 ng/mL:
52%, 5 ng/mL: 62%, 10 ng/mL: 76%, 5 µg/mL: 79%; 10 µg/mL: 60%) compared to day 0 (37%). The culture of ovarian tissue in the presence
of 10 ng/mL or 5 µg/mL increased the percentage of developing follicles when compared to other cultivated treatments (P