Thymidine monolayer cultures

Cytotoxic effect. Gas phase of mainstream cigarette smoke, in monolayer culture of mouse lung epithelial cells, produced an increase in cytotoxicity in a dose-dependent manner. Cell viability of cultures exposed to gas phase with only the nonorganic components was equivalent to controls. Removal of volatile organic constituents resulted in almost elimination of cytotoxicity of the smoke . Smoke condensate and tobacco extract, at high concentrations in Lewis lung adenocarcinoma cells and mice spleen lymphocytes, were cytotoxic. Smaller doses increased thymidine incorporation in both cell types. Lymphocytes were more susceptible to the toxic effect of tobacco prod-... 

Fig. 14.1. Time course of SV40 infection. Monkey cells in monolayer culture, infected with SV40 at 1-10 p.f.u. per cell. T antigen may be detected by immunofluorescence, viral DNA synthesis by labelling with [yH]thymidine followed by separation of viral DNA (by Hirt extraction, SDS gradient centrifugation or agarose gel electrophoresis) and mature virions by infectivity using a plaque assay. (Data from Tooze, 1973 Girard et al., 1975 and Basilico and Zouzias, 1976.)...

Suspension cultures can be achieved using F12 medium with 10% foetal bovine serum (FBS) and gassing the culture vessels continuously with 5% CO2. The doubling time under these conditions is typically 18 h versus 14 h for monolayer cultures. The growth requirements of CHO dhfr cells are hypoxanthine (or adenine), glycine and thymidine in addition to the usual requirement of CHO cells for proline. 

Production of Mucosal Damage 2.3.1.2.1 Cell culture Stimulated neutrophils are known to be cytotoxic to cells in vitro (Dull et al., 1987 Dallegri et al., 1990 Grisham et al., 1990b). Several in vitro systems have been used to demonstrate oxidative damage to intestinal cells. Xanthine/XO increased Cr release and decreased [ H]thymidine uptake by IEC-18 small intestinal epithelial cell monolayers in a dose-dependent manner (Ma et al., 1991). Rat enterocytes show decreased trypan blue exclusion and increased protein release when incubated with neutrophils stimulated... 

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