Ali Atef, Coenen Karine, Bousquet Daniel, Sirard Marc-André
Department of Animal Science, Centre de Recherche en Biologie de la Reproduction, Laval University, Ste-Foy, Que., Canada G1K 7P4.
Theriogenology. 2004 Dec;62(9):1596-606. doi: 10.1016/j.theriogenology.2004.03.011.
Protein supplementation during in vitro maturation can profoundly affect both the rate and overall efficiency of the maturation procedure. The present study was conducted to assess the ability of different concentrations (1, 5, and 10%) of bovine follicular fluid (bFF) to support in vitro maturation of oocytes and subsequent developmental capacity. The bFF was derived either from competent follicles ( > 8 mm) obtained by transvaginal recovery following superovulation or from a pool of small follicles (2-5 mm) from abbatoir-derived ovaries. Bovine oocytes were cultured for 24 h in synthetic oviduct fluid medium (m-SOF) supplemented with polyvinylpyrrolidone. Following fertilization and embryo culture, more oocytes (P < 0.05) reached the blastocyst stage when oocytes were cultured with 5% bFF from competent follicles (41 +/- 3.7%) compared with bFF derived from small follicles (16 +/- 2.9%). Estradiol and recombinant human follicle stimulating hormone added to the competent bFF during maturation acted in synergy to increase blastocyst production rate (P < 0.05); this blastocyst production rate (57 +/- 1.2%) was higher than those obtained with the addition of these two hormones to bFF derived from small follicles (26 +/- 2.9%). The quality of blastocysts obtained was reflected by inner cell mass (51.30 +/- 3.5 and 25.50 +/- 3.7) and trophectoderm cell numbers (99.72 +/- 2.5 and 94.80 +/- 4.7) for bFF from competent and small follicles, respectively. In conclusion, follicular fluid originating from competent follicles increased the developmental competence of abbatoir-derived oocytes.
体外成熟培养过程中的蛋白质补充可深刻影响成熟程序的速率和整体效率。本研究旨在评估不同浓度(1%、5%和10%)的牛卵泡液(bFF)支持卵母细胞体外成熟及后续发育能力的情况。bFF来源于超排后经阴道回收获得的优势卵泡(> 8 mm)或来自屠宰场卵巢的小卵泡池(2 - 5 mm)。牛卵母细胞在添加聚乙烯吡咯烷酮的合成输卵管液培养基(m - SOF)中培养24小时。受精和胚胎培养后,与来自小卵泡的bFF(16% ± 2.9%)相比,用来自优势卵泡的5% bFF培养卵母细胞时,更多卵母细胞(P < 0.05)发育到囊胚阶段(41% ± 3.7%)。成熟培养期间向优势bFF中添加雌二醇和重组人促卵泡激素具有协同作用,可提高囊胚生成率(P < 0.05);该囊胚生成率(57% ± 1.2%)高于向来自小卵泡的bFF中添加这两种激素时的囊胚生成率(26% ± 2.9%)。对于来自优势卵泡和小卵泡的bFF,所获得囊胚的质量分别以内细胞团数量(51.30 ± 3.5和25.50 ± 3.7)和滋养外胚层细胞数量(99.72 ± 2.5和94.80 ± 4.7)来反映。总之,来自优势卵泡的卵泡液提高了屠宰场来源卵母细胞的发育能力。