【摘要】 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(PGC s)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O ct-4转录因子;(2)收集妊娠29~100 d牛胎儿28例,从生殖嵴(腺)或类似物中分离克隆水牛PGC s,23例出现PGC s细胞集落。其中胎龄小于45 d(<3.0 cm)和45~55 d(3.0~5.0 cm)各7例,全部出现PGC s细胞集落;55~70 d(5.0~9.0 cm)9例,有7例出现PGC s细胞集落,大于70 d的5例,有2例出现出现PGC s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55 d的水牛胎儿易于分离培养形成PGC s集落。更多还原

【Abstract】 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,multilayered 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 29～100 d,7 clones were from 7 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.更多还原