Gliedt D W, Rosenkrans C F, Rorie R W, Munyon A L, Pierson J N, Miller G F, Rakes J M
Department of Animal Science, University of Arkansas, Fayetteville 72701, USA.
J Dairy Sci. 1996 Apr;79(4):536-42. doi: 10.3168/jds.S0022-0302(96)76397-X.
Our objective was to optimize in vitro maturation conditions of bovine oocytes as assessed by embryo development. In Experiment 1, cumulus-oocyte complexes were matured in either M-199 or RPMI-1640. Each medium was supplemented with an antibiotic-antimycotic solution (1%) and estrous cow serum (20%). Cumulus cell expansion after 24 h was greatest for cumulus-oocyte complexes matured in RPMI-1640. Morulae development on d 7 was greater (21.1%) for oocytes matured in M-199 than for oocytes that matured in RPMI-1640 (9.6%). In Experiment 2, cumulus-oocyte complexes were matured in M-199 supplemented with antibiotic-antimycotic solution (1%). Main effects were serum type (20%; estrous cow serum vs. superstimulated estrous cow serum) and coculture (with or without bovine oviductal epithelial cells). The percentage of oocytes developing into blastocysts (d 9) was higher for oocytes matured in estrous cow serum regardless of coculture. In Experiment 3, effects of estradiol-17 beta (0, 1, and 2 micrograms/ml) and equine LH (0, 10, 20, and 30 micrograms/ml) on cumulus cell expansion and development after fertilization were determined. Cumulus cell expansion and blastocyst development decreased with estradiol-17 beta in the maturation medium, but LH in the medium enhanced expansion of cumulus cells and blastocyst development.
我们的目标是优化牛卵母细胞的体外成熟条件，并通过胚胎发育进行评估。在实验1中，卵丘-卵母细胞复合体在M-199或RPMI-1640中成熟。每种培养基均添加了抗生素-抗真菌溶液（1%）和发情母牛血清（20%）。在RPMI-1640中成熟的卵丘-卵母细胞复合体在24小时后卵丘细胞扩张最大。在第7天，在M-199中成熟的卵母细胞的桑葚胚发育率（21.1%）高于在RPMI-1640中成熟的卵母细胞（9.6%）。在实验2中，卵丘-卵母细胞复合体在添加了抗生素-抗真菌溶液（1%）的M-199中成熟。主要因素是血清类型（20%；发情母牛血清与超刺激发情母牛血清）和共培养（有无牛输卵管上皮细胞）。无论是否共培养，在发情母牛血清中成熟的卵母细胞发育成囊胚（第9天）的百分比更高。在实验3中，测定了17β-雌二醇（0、1和2微克/毫升）和马促黄体素（0、10、20和30微克/毫升）对卵丘细胞扩张和受精后发育的影响。成熟培养基中的17β-雌二醇会降低卵丘细胞扩张和囊胚发育，但培养基中的促黄体素会增强卵丘细胞扩张和囊胚发育。
