大鼠原始生殖细胞的分离培养

Isolation and Culture of Rat Primordial Germ Cells

以不同日龄的大鼠胎仔为材料，对其生殖巢进行解剖分离，并作形态观察．结果发现随着日龄的增加，生殖巢由长变短，由薄增厚，并在１４．５ｄ的生殖巢上发现了明显的性分化．对不同日龄的生殖巢用ＥＤＴＡ－Ｔｒｙｐｓｉｎ在３７℃下进行解离，可以得到原始生殖细胞（ＰＧＣｓ），较早日龄的ＰＧＣｓ形态较不规则，有大量的伪足伸出，运动能力较强．随着日龄的增大，ＰＧＣｓ的形态有所变化，椭圆形、圆形居多，并且运动能力亦有所下降．ＰＧＣｓ的体积比一般的体细胞大，细胞核多为圆形，体积硕大，有多个核仁．将消化液处理后的产物进行培养，ＰＧＣｓ一般能在含血清的ＤＭＥＭ培养液中悬浮生存一周左右，在此期间也有部分ＰＧＣｓ发生皱缩死亡，大量的体细胞在皿底成纤维化，血细胞逐渐死亡．

The morphology of rat gonads obtained from both sexes 12.5,13.5,and 14.5 days post coitum(dpc) has been examined. Results showed that fetal rat gonads turned short and thick with the increasing of dpc.The sexualdifferentiation was very significant in 14.5dpc gonads of rat fetal.Gonadswere treated by EDTA and Trypsin in 37 ℃,then they were disaggregation.The PGCs (primordial germ cells) obtained from early dpc gonads had irregular morphology ,and they can make active amoeboid movement,which had been demonstrated by timelapse photography.With the increasing of dpc,PGCs tured round and regular,the locomotory activity decreased.PGCs has a clear appearence and can be recognized from most somatic cells.Its nuclei is spherical and large, and has several prominent nucleoli.PGCs can be cultured for at least one week in DMEM culture media with fetal calf serum .The divison of PGCs failed to be examined. Most of somatic cell differentiat fibroblast.