Sakurai H, Sakurai H, Tegoshi T, Takagi T, Saito A, Suzuki N, Seitz H M
Department of Veterinary Physiology, Obihiro University, Hokkaido, Japan.
Zentralbl Bakteriol Mikrobiol Hyg A. 1987 May;264(3-4):468-77. doi: 10.1016/s0176-6724(87)80070-6.
To establish a method of mass and long-term propagation of Toxoplasma in suspension cultures and to find a technique for separating the organisms from host cells, basic investigations were carried out in a microcomputer controlled system. By using an arbitrary unit "the medium index" host HeLa cells were continuously supplied to Toxoplasma organisms for about 30 days, at cycles of 4 days. HeLa cells were well infected with Toxoplasma in the suspension cultures by leaving the medium unstirred for 15 min intervals. The separation of Toxoplasma organisms from cells was improved by treating toxoplasmas and HeLa cells mixtures with 0.4% trypsin. The total number of separated organisms obtained from 29 days cultivation period was 4.9 X 10(9). The mean generation time of Toxoplasma organisms on the 0, 10, 20 and 30 day's cultivation was calculated approximately 7.4, 9.4, 9.3 and 7.8 h, respectively.
为建立弓形虫在悬浮培养中的大规模长期繁殖方法,并找到从宿主细胞中分离该生物体的技术,在微机控制系统中进行了基础研究。通过使用任意单位“培养基指数”,以4天为周期,连续约30天向弓形虫生物体供应宿主HeLa细胞。通过每隔15分钟不搅拌培养基,HeLa细胞在悬浮培养中被弓形虫良好感染。用0.4%胰蛋白酶处理弓形虫与HeLa细胞的混合物,可改善从细胞中分离弓形虫生物体的效果。从29天培养期获得的分离生物体总数为4.9×10⁹。在培养的第0、10、20和30天,弓形虫生物体的平均世代时间分别约为7.4、9.4、9.3和7.8小时。