Wu Liang, Chen Sheng-xia, Li Lin-jie, Che Fei-hu, Deng Hong-yan, Jiang Xu-gan
School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212013, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2008 Dec 30;26(6):452-6.
To establish a stable and efficient in vitro culture model for tachyzoites of Toxoplasma gondii RH strain.
Tachyzoites were inoculated into HeLa cells to establish an in vitro culture system. The proliferation of tachyzoites was observed under microscope by the method of Giemsa stain. At the same time, the longterm tachyzoites maintenance in HeLa cells was established, and the effect of different temperature and time on the yield and motility of tachyzoites were observed.
The RH strain tachyzoites were cultured and maintained in HeLa cells. Most HeLa cells were destroyed 96 h after inoculation. In the long-term culture system, the proliferation of tachyzoites was stable and its virulence to mouse showed no decrease. Furthermore, tachyzoites in this system proliferated by 5-20 times and (1-5) x 10(7) tachyzoites were harvested. When cultured in HeLa cells at 37 degrees C for 72h then at 25 degrees C for another 120 h, the tachyzoites proliferated by more than 40 times with a motility rate of over 90%. However, rare HeLa cells left in the medium were found.
Tachyzoites of T. gondii RH strain can be subcultured in HeLa cells for a long time, and high proliferation rate of tachyzoites can be obtained from this in vitro culture system.
建立稳定、高效的刚地弓形虫RH株速殖子体外培养模型。
将速殖子接种于HeLa细胞，建立体外培养体系。采用姬姆萨染色法在显微镜下观察速殖子的增殖情况。同时，建立HeLa细胞中长期维持速殖子的方法，观察不同温度和时间对速殖子产量和活力的影响。
RH株速殖子可在HeLa细胞中培养并维持。接种后96 h多数HeLa细胞被破坏。在长期培养体系中，速殖子增殖稳定，对小鼠的毒力未降低。此外，该体系中速殖子增殖5 - 20倍，收获(1 - 5)×10(7)个速殖子。当在37℃下于HeLa细胞中培养72 h，然后在25℃下再培养120 h时，速殖子增殖超过40倍，活力率超过90%。然而，培养基中剩余的HeLa细胞很少。
刚地弓形虫RH株速殖子可在HeLa细胞中长时间传代培养，且该体外培养体系可获得高增殖率的速殖子。
