水牛原始生殖细胞(PGCs)的分离培养及生物学特性的研究

Studies on isolation,culture and biological charactristics of buffalo primordial germ cells

以水牛胎儿为材料，从生殖腺中分离培养出牛原始生殖细胞，并对水牛原始生殖细胞的分离与克隆的影响因素进行了探讨。结果发现：（1）分离培养出的原生殖细胞（PGCs）呈集落状生长．细胞堆积密集，细胞之间界限不清．细胞与周围的成纤维细胞界限明显；未分化的细胞表达碱性磷酸酶（AKP）以及Oct-4转录因子；（2）收集妊娠29～100d牛胎3L28例，从生殖嵴（腺）或类似物中分离克隆水牛PGCs，23例出现PGCs细胞集落。其中胎龄小于45d（〈3．0cm）和45～55d（3．0～5．ocm）各7例，全部出现PGCs细胞集落；55～70d（5．0～9．0cm）9例，有7例出现PGCs细胞集落，大于70d的5例，有2例出现出现PGCs细胞集落。结果表明，水牛原生殖细胞具有多能性特性，胎龄小于55d的水牛胎儿易于分离培养形成PGCs集落。

The major objectives of this study are to isolate and culture primordial germ cells （PGCs） from buffalo gonad as well as to demonstrate the pluripotent of the cells. The factors influenced the efficiency of the isolation and culture buffalo PGCs were also studied. The PGCs isolated from buffalo fetuses grew in large, muhilayered colonies which were densely packed with an obvious border, and expressed AKP, Oct-4. Among 23 PGCs clones derived from 28 normal buffalo fetuses of 29～100d, 7 clones were from7 fetuses less than 45 d, 7 clones were from 7 fetuses of 45 ～55 d, 7 clones were from 9 fetuses of 55～70 d, and 2 clones were from 5 fetuses of more than 70 d. These results indicate that buffalo PGCs have typical pluripotent characteristics, and PGCs isolated from fetuses less than 55 d are more efficient to form clones.