In vitro maturation supplements affect developmental competence of bovine cumulus-oocyte complexes and embryo quality after vitrification |
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Authors: | Räty Mervi Ketoja Elise Pitkänen Timo Ahola Virpi Kananen Kirsi Peippo Jaana |
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Affiliation: | aMTT Agrifood Research Finland, Biotechnology and Food Research, FI-31600 Jokioinen, Finland;bUniversity of Kuopio, Department of Biosciences, FI-70211 Kuopio, Finland |
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Abstract: | Oocyte quality affects subsequent embryo development and quality. We examined the impact of bovine oocyte in vitro maturation (IVM) conditions on subsequent embryo yield, quality and cryosurvival. Cumulus–oocyte complexes (COCs) were sampled for cytological and gene expression analysis after IVM in TCM199 supplemented with 10% fetal bovine serum (FBS), 4 mg/ml of fatty-acid-free bovine serum albumin (FAFBSA), 4 mg/ml of polyvinylpyrrolidone (PVP), FAFBSA with epidermal growth factor (EGF, 100 ng/ml) and insulin-like growth factor 1 (IGF-I, 100 ng/ml) (FAFBSAGF), PVP with EGF and IGF-I (PVPGF) or PVP with single strength BME and MEM amino acids (PVPAA). The remaining COCs were fertilized. On day 7 (IVF = day 0) quality 1 blastocysts were vitrified or analyzed for glucose transporter 1 (Glut-1) expression levels. The remaining blastocysts (days 7–9) were evaluated for morphology and total cell counts. After warming, survival and hatching rates were evaluated followed by total cell counts and Glut-1 expression levels. Only PVPGF IVM resulted in embryo production rates comparable to those recorded with FBS IVM. Growth factors with FAFBSA and amino acids with PVP reduced embryo production rates whereas the effect of the growth factors with PVP was negligible. Insulin-like growth factor 2 binding protein 3 (IGF2BP3) and beta cell translocation gene 4 (BTG4) were revealed as potential candidates for oocyte developmental competence, and secreted protein, acidic and rich in cysteine (SPARC) for cumulus cell expansion. There were no differences among treatments in hatching rates of vitrified embryos after warming. However, total cell numbers and Glut-1 expression levels at 72 h were affected. |
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Keywords: | Amino acid Epidermal growth factor Fatty-acid-free bovine serum albumin Gene expression Glucose transporter 1 Insulin-like growth factor 1 Open pulled straw Polyvinylpyrrolidone Serum |
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