By Peter Malcolm Wallis, Brian R. Hammond
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The supernatants (cell free extract) were used to assay the cytotoxicity. Fresh TPS1 complete medium without the parasite and minimal essential medium (MEM) with 5% of calf serum were used as controls. 5 × 105 cells/mL to initiate the cultures for cellparasite interactions. For this purpose, effects of inocula of varying number of trophozoites ranging from 1 × 104 to 1 × 107 on HeLa and Vero cells at different intervals (0, 5, 18, 24, 48, 72 and 96 h) after incubation at 37°C were studied. 5 × 105 cells/mL • Corresponding author.
Cell free culture supernatant was collected from 48 h old cultures after chilling and centrifugation as described above. After washing with phosphate buffered saline, trophozoites were ruptured by a freeze/thaw cycle and then centrifuged (105 × g for 1 h) at 4°C. The supernatants (cell free extract) were used to assay the cytotoxicity. Fresh TPS1 complete medium without the parasite and minimal essential medium (MEM) with 5% of calf serum were used as controls. 5 × 105 cells/mL to initiate the cultures for cellparasite interactions.
33:11791183. Microbiol. 45:6574. 80:4952. Parasitologia 4:510514. 12:851853. Science 92:416417. A. Cysts formed in vitro were both morphologically and immunologically similar to Giardia cysts formed in vivo and also were viable, as demonstrated by the uptake of fluorogenic dyes. The inability of Giardia trophozoites to undergo encystation in vitro has raised questions as to whether host related factors, such as gutassociated microorganisms or nutritive factors and stimuli from the small intestine, might play a role in this process.