Lipid content and apoptosis of in vitro-produced bovine embryos as determinants of susceptibility to vitrification

Autor(es)

Sudano, Mateus Jose ^[1] ; Paschoal, Daniela Martins ^[1] ; Rascado, Tatiana da Silva ^[1] ; Ona Magalhaes, Luis Carlos ^[1] ; Crocomo, Leticia Ferrari ^[1] ; de Lima-Neto, Joao Ferreira ^[1] ; Landim-Alvarenga, Fernando da Cruz ^[1]

Afiliação do(s) autor(es)

^[1] UNESP, Sao Paulo State Univ, Sch Vet Med & Anim Sci, FMVZ, Dept Anim Reprod & Vet Radiol, BR-18618970 Botucatu, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Theriogenology; v. 75, n. 7, p. 1211-1220, APR 15 2011.
• Citações Web of Science: 64

Resumo

The objective was to evaluate supplementation of fetal calf serum (FCS) and phenazine ethosulfate (PES), a metabolic regulator that inhibits fatty acid synthesis, in culture media during in vitro production (IVP) of bovine embryos. Taking oocyte fertilization (n = 4,320) as Day 0, four concentrations of FCS (0, 2.5, 5, and 10%) and three periods of exposure to PES (without addition-Control; after 60 h-PES Day 2.5 of embryo culture; and after 96 h-PES Day 4) were evaluated. Increasing FCS concentration in the culture media enhanced lipid accumulation (P < 0.05), increased apoptosis in fresh (2.5%: 19.1 +/- 1.8 vs 10%: 28.4 +/- 2.3, P < 0.05; mean +/- SEM) and vitrified (2.5%: 42.8 +/- 2.7 vs 10%: 69.2 +/- 3.4, P < 0.05) blastocysts, and reduced blastocoele re-expansion after vitrification (2.5%: 81.6 +/- 2.5 vs 10%: 67.3 +/- 3.5, P < 0.05). The addition of PES in culture media, either from Days 2.5 or 4, reduced lipid accumulation (P < 0.05) and increased blastocoele re-expansion after vitrification (Control: 72.0 +/- 3.0 vs PES Day 2.5: 79.9 +/- 2.8 or PES Day 4: 86.2 +/- 2.4, P < 0.05). However, just the use of PES from D4 reduced apoptosis in vitrified blastocysts (Control: 52.0 +/- 3.0 vs PES Day 4: 39.2 +/- 2.4, P < 0.05). Independent of FCS withdrawal or PES addition to culture media, the in vivo control group had lesser lipid accumulation, a lower apoptosis rate, and greater cryotolerance (P < 0.05). The increased lipid content was moderately correlated with apoptosis in vitrified blastocysts (r = 0.64, P = 0.01). In contrast, the increased apoptosis in fresh blastocysts was strongly correlated with apoptosis in vitrified blastocysts (r = 0.94, P < 0.0001). Therefore, using only 2.5% FCS and the addition of PES from Day 4, increased the survival of IVP embryos after vitrification. Moreover, embryo quality, represented by the fresh apoptosis rate, was better than lipid content for predicting embryo survival after vitrification. (C) 2011 Elsevier Inc. All rights reserved. (AU)

• Processo FAPESP: 07/57766-4 - Vitrificação de embriões bovinos produzidos in vitro após suplementação do meio de cultivo embrionário com phenazine ethosulfate e soro fetal bovino no período de pós compactação embrionária
• Beneficiário: Mateus José Sudano
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 08/51378-5 - Uso de forskolin ou phenazine ethosulfate visando a diminuicao do teor de lipideos intracelulares em embrioes ovinos produzidos in vitro destinados a vitrificacao.
• Beneficiário: Fernanda da Cruz Landim
• Modalidade de apoio: Auxílio à Pesquisa - Regular