Eppig J J
J Exp Zool. 1976 Dec;198(3):375-82. doi: 10.1002/jez.1401980311.
A method is described for the dissociation of mouse ovaries and the isolation of oocytes free of somatic cells by agitating pieces of ovary in collagenase and deoxyribonuclease in a calcium and magnesium free salt solution. This method yielded about 50% of the growing oocytes from immature mice. The utilization of exogenously administered 14C-labelled energy sources by oocytes in various growth stages was determined by measurement of evolved 14CO2. Little or no evolution of 14CO2 was detected from oocytes of any size incubated in 14C-glucose, lactate or succinate. The production of 14CO2 from 14C-pyruvate increased logarithmically when plotted against increasing oocyte volume with a plateau occurring after occytes reached a volume of 65,500 mum3 (50 mum diameter). Thus, the pattern of energy metabolism for oocyte maturation and early egg cleavage, wherein glucose and lactate are not utilized as efficiently as pyruvate, has been established by the earliest stages of oocyte growth.
本文描述了一种分离小鼠卵巢并通过在无钙镁盐溶液中用胶原酶和脱氧核糖核酸酶搅拌卵巢组织来分离无体细胞卵母细胞的方法。该方法从未成熟小鼠中获得了约50%的生长中卵母细胞。通过测量释放的14CO2来确定不同生长阶段的卵母细胞对外源给予的14C标记能量源的利用情况。在14C -葡萄糖、乳酸盐或琥珀酸盐中培养的任何大小的卵母细胞几乎都未检测到14CO2的释放。当以增加的卵母细胞体积作图时，14C -丙酮酸产生的14CO2呈对数增加，当卵母细胞达到65,500立方微米（直径50微米）体积后出现平台期。因此，在卵母细胞生长的最早阶段就已确立了卵母细胞成熟和早期卵裂的能量代谢模式，其中葡萄糖和乳酸盐的利用效率不如丙酮酸。
