MTT Agrifood Research Finland, Biotechnology and Food Research, Jokioinen, Finland.
Cryobiology. 2011 Dec;63(3):245-55. doi: 10.1016/j.cryobiol.2011.09.134. Epub 2011 Oct 1.
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.
卵母细胞质量影响随后的胚胎发育和质量。我们研究了牛卵母细胞体外成熟（IVM）条件对随后的胚胎产量、质量和冷冻保存的影响。在添加 10%胎牛血清（FBS）、4mg/ml 无脂肪酸牛血清白蛋白（FAFBSA）、4mg/ml 聚乙烯吡咯烷酮（PVP）、含表皮生长因子（EGF，100ng/ml）和胰岛素样生长因子 1（IGF-I，100ng/ml）的 TCM199 中进行 IVM 后，对卵丘-卵母细胞复合物（COC）进行细胞学和基因表达分析（FAFBSAGF）、含 EGF 和 IGF-I 的 PVP（PVPGF）或含单倍 BME 和 MEM 氨基酸的 PVP（PVPAA）。其余 COC 进行受精。在第 7 天（IVF=第 0 天），对质量 1 囊胚进行玻璃化或葡萄糖转运蛋白 1（Glut-1）表达水平分析。剩余的囊胚（第 7-9 天）进行形态学和总细胞计数评估。解冻后，评估存活率和孵化率，然后进行总细胞计数和 Glut-1 表达水平分析。只有 PVPGF IVM 导致胚胎产率与 FBS IVM 记录的产率相当。含有 FAFBSA 的生长因子和含有 PVP 的氨基酸降低了胚胎产率，而 PVP 中的生长因子的影响可以忽略不计。胰岛素样生长因子 2 结合蛋白 3（IGF2BP3）和β细胞易位基因 4（BTG4）被揭示为卵母细胞发育能力的潜在候选基因，而分泌蛋白、酸性富含半胱氨酸（SPARC）则用于卵丘细胞扩张。在解冻后的玻璃化胚胎孵化率方面，不同处理组之间没有差异。然而，总细胞数和 72 小时的 Glut-1 表达水平受到影响。
