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Comparative susceptibility of different cell lines for culture of Toxoplasma gondii in vitro
B K Park,*1H R Moon,2J R Yu,3J Kook,4J Y Chai,4 and S H Lee4
Department of Pediatrics, Gyeongsang Institute of Cancer Research, Gyeongsang National University, Chinju 660-280, Korea.
Received June 24, 1993; Accepted August 06, 1993.
Abstract
In order to establish a useful cell culture system for T. gondii, we compared the degree of proliferation of T. gondii tachyzoites among 8 different cell lines; 2 kinds of normal animal cells (MDCK-canine kidney cells; Vero-monkey kidney cells) and 6 kinds of human tumor cells (A 549, PC 14-lung cancer cells; SNU 1, SNU 16, MKN 45-stomach cancer cells; HL-60-promyelocytic leukemia cells), through morphological observation and 3H-uracil uptake assay. The degree of susceptibility to infection with T. gondii tachyzoites was highest in A 549 and PC 14 cells, medium in Vero, HL-60, MDCK and SNU 1, and lowest in SNU 16 and MKN 45 cells. The kinetics of T. gondii multiplication during the post-infection 60 hours were highly dependent upon the dose of tachyzoites administered and the duration of cultivation. These results show that A 549 and PC 14 are the most suitable cell lines among the 8 tested for the growth and multiplication of T. gondii in vitro.
Figures
Figs. 1-6 Fig. 1. An A 549 cell (a highly susceptible cell line) infected with 10 × 105/ml tachyzoites of T. gondii, 12 hrs after infection. Note inrtacellular endodyogeny (arrows) of tachyzoites. Modified Giemsa, × 1,000. Fig. 2. A 549 cells infected with 10 × 105/ml tachyzoites of T. gondii, 12 hrs after infection. Note that the cytoplasm of two cells is being ruptured due to proliferation of tachyzoites. Modified Giemsa, ×1,000. Fig. 3. Vero cells (a medium susceptible cell line) infected with T. gondii 12 hrs previously. Note two just divided tachyzoites (arrow) and circularly arranged ones (arrow) in the host cell cytoplasm. Also note a Vero cell which is about to rupture due to many tachyzoites in the cytoplasm. Modified Giemsa, ×1,000. Fig. 4. A Vero cell infected with T. gondii, 12 hrs previously. Numerous tachyzoites are arranged in a rosette form and occupying the whole cytoplasm of a dividing host cell. Modified Giemsa, ×1,000. Fig. 5. Smear from culture of PC 14 cells (a highly susceptile cell line) infected with T. gondii, 12 hrs previously. Tachyzoites are bing liberated from a cell (arrows). Freed tachyzoites are also seem from the culture fluid. Modified Giemsa, ×1,000. Fig. 6. MKN 45 cells (a less susceptible cell line) infected with T. gondii, 12 hus previously. Note intracellular parasites (arrows) and liberated ones. The intracellular ones, however, look not so good in their condition morphologically. Modified Giemsa, ×1,000.
Fig. 7 3H-uracil uptake of T. gondii cultured in various cell lines according to the cultivation time (12 hrs, 36 hrs and 60 hrs). Cultures were inoculated with 2 × 105/ml tachyzoites. A 549 and PC 14 cells show highest uracil uptake by parasites up to 60 hrs of incubation compared to other cell lines.
Fig. 8 3H-uracil uptake of T. gondii cultured in various cell lines according to the cultivation time (12 hrs, 36 hrs and 60 hrs). Cultures were inoculated with 50 × 105/ml tachyzoites. Uracil uptake declines after 12 hrs in most of the cell lines. A 549, PC 14 and Vero cells show highest uracil uptake by parasites.
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