Long-term in vitro suspension culture of Toxoplasma and HeLa cells.
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.
Sakurai
H
H
Department of Veterinary Physiology, Obihiro University, Hokkaido, Japan.
Sakurai
H
H
Tegoshi
T
T
Takagi
T
T
Saito
A
A
Suzuki
N
N
Seitz
H M
HM
D000818:Animals; D002470:Cell Survival; D003470:Culture Media; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000469:parasitology; D008838:Microcomputers; D013997:Time Factors; D014122:Toxoplasma / Q000254:growth & development*; D014357:Trypsin
D003470:Culture Media; D014357:Trypsin