细粒棘球蚴生发层细胞的体外培养

Studies on the Continual Cultivation of Germinal Cells from Hydatid Cysts of Echinococcus granulosus

将分散的绵羊源细粒棘球蚴生发层细胞在５％ＣＯ２、３７℃恒温箱中进行连续传代培养，细胞培养液为ＲＰＭＩ１６４０完全培养基，细胞支持物在１～１４代时交替为２４孔细胞培养板和用胶原蛋白包被的２４孔细胞培养板，１４代后仅为２４孔细胞培养板。细胞经４０～４５ｄ传代培养（８代）后增殖缓慢。１２代开始至２５代止，各孔细胞又逐步转变成快速增殖细胞，第２６代细胞在对数增长期内的倍增时间约为２４ｈ。这一细胞增殖速率持续至３４代后约有减缓。传代增殖细胞呈小点形、圆形和椭圆形，大部分行悬浮生长。行贴壁生长的细胞经长期培养后融合成合胞体。

The germinal cells obtained from echinococcal cysts in the feral sheep have been cultivated continually. The medium was RPMI1640 with 10% 20% fetal calf serum. The cell supports were 24 well cell culture plate and collagen coated ones alternately during the first few subcultures, and only 24 well cell culture plate after passage 14. Subcultivation was carried out at the split up rate of 1∶2 1∶3 every 5 7 days up to passage 8(40 45 days). Afterwards, the growth rate decreased so that subcultivation was done every 5 7 days at the rate of 1∶1 1∶1.5 splits. From passage 12(91 days) to passage 25 (161 days), the multiplication rate of cells in respective wells increased, so, the cells in a well could be split into three or four wells every 3 4 days. Such a high multiplication continued to passage 34 (216 days), and then the subcultured cells grew in a slightly lower rate, which subcultivation could be done in 1∶2 1∶3 splits every 7 days. Population doubling time in the logarithmic phase of growth at passage 26 was approximately 24 hours. The subcultured cells showed circular and elliptical in shape and with smooth surface examined by light microscopy. Most of the cells were suspending, wherease the others sticking which could fuse into syncytia and become tissue like masses eventually.