屠宰绵羊卵巢卵母细胞的体外培养

THE MATURATION OF SIAUGHTERED OVINE OOCYTES CULTURED IN VITRO

为了使屠宰绵羊卵巢卵母细胞能够用于体外受精,本文着重探索了使卵巢卵母细胞体外培养成熟的方法和条件。实验结果表明,以TCM-199加10％FCS作为基本培养基,培养24～25小时,可以使绵羊卵巢卵母细胞培养成熟,其成熟率可达55.5％(435/784)。如果在培养液内添加hCG(0.02mg/ml)或LH(0.01mg/ml),并且尽可能保持卵丘细胞的完整,则可以使成熟率提高到76.9％(140/182)～82.9％(112/135)。

In order to provide a large number of oocytes for fertilization in vitro, an effective culturing method were developed which was found suitable for large scale preparation of matured oocytes. There were 784 oocytes collected from the Graafian follicles of newly slaughtered sheep have been grouped into three types: Type A , the oocytes surrounded by cumulus cells; Type B, the oocytes surrounded partly by cumulus cells; and Type C, denuded oocytes. Each type of oocytes have been cultured in three different kinds of medium. 1 ), M199 + FCS; 2), Ml99 + FCS + hCG; 3), M199 + FCS + LH, at 37@@ under water-sa tured atmosphere of 95% air-5% CO2 fpr 24~28 hours. After fixation and staining, the percentage of oocytes maturation can be calculated under the phase-contrast microscope. Out of a total number of 784 cultured oocytes, 435 (55.5% ) matured oocytes were obtained. The maturation rate of Type A were 76.9% and 82.9% with LH and hCG respectively. It was demonstrated that in M199 containing PCS , oocytes are able to mature. But a higher maturation rate requires addition of LH or hCG.