IN-VITRO GROWTH AND MATURATION OF PIG OOCYTES
IN-VITRO GROWTH AND MATURATION OF PIG OOCYTES
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DOI：
10.1530/jrf.0.1000333
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发表时间：
1994-03-01
期刊：
影响因子：
--
通讯作者：
KATO, S
中科院分区：
文献类型：
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作者：
HIRAO, Y;NAGAI, T;KATO, S
从猪卵巢中分离出含有直径为70-89.5 μ m卵母细胞的腔前卵泡，并在血清、FSH和雌二醇存在下，在胶原凝胶中培养长达16天。随着培养的进行，卵泡腔逐渐形成。卵母细胞被颗粒细胞包围，卵母细胞和包围的卵丘细胞的突起之间的接触在培养期间得以维持。培养16天后，30-40%的卵母细胞外观正常，约半数卵母细胞直径大于100 μ m。当体外培养的卵母细胞从卵泡中释放出来，并在改良的Krebs-Ringer碳酸氢盐溶液中再培养48小时时，分别有6%、30%和60%的大于90、100和110 μ m的卵母细胞发生生殖囊泡破裂。在直径超过110 μ m的卵母细胞中观察到40%的卵母细胞进入中期II，而直径小于90 μ m的卵母细胞没有恢复减数分裂。卵母细胞的大小和减数分裂能力之间的关系是相似的卵母细胞在体外或体内生长。体外培养成熟的卵母细胞被精子穿透并形成雌性原核，但精子头部的解凝不完全。本研究首次证明猪腔前卵泡卵母细胞在体外能够发育成熟，获得减数分裂能力，并能被精子穿透。
Preantral follicles containing oocytes of 70-89.5 mu m in diameter were isolated from pig ovaries and cultured in collagen gel for up to 16 days, in the presence of serum, FSH and oestradiol. Formation of follicular antra occurred as the culture proceeded. The oocytes had been enclosed by granulosa cells and contacts between the oocytes and processes of the enclosing cumulus cells were maintained over the culture period. After 16 days of culture, 30-40% of the oocytes were of normal appearance, and the diameters of about half of these oocytes were larger than 100 mu m. When the oocytes grown in vitro were liberated from the follicles and cultured for a further 48 h in modified Krebs-Ringer bicarbonate solution, 6, 30 and 60% of the oocytes larger than 90, 100 and 110 mu m underwent germinal vesicle breakdown, respectively. Progression to metaphase II was observed in 40% of oocytes that were over 110 mu m in diameter, whereas no oocyte less than 90 mu m in diameter resumed meiosis. The relationship between the size and meiotic competence of oocytes was similar for oocytes grown in vitro or in vivo. Oocytes grown and matured in vitro were penetrated by spermatozoa and formed a female pronucleus, but decondensation of the sperm head was incomplete. The results demonstrate for the first time that pig oocytes from preantral follicles can grow up to their final size, acquire meiotic competence, and be penetrated by spermatozoa in vitro.