Macrophage culture: influence of species-specific incubation temperature

Cultured mammalian cells are traditionally maintained at 37 degrees C, despite the fact that core body temperatures differ considerably among mammals. Considering the body temperature of the adult pig, comparison was made of porcine macrophage cultures maintained at 37 degrees C and 39.2 degrees C. Examination of the cells showed that granularity was higher in macrophages maintained at 39.2 degrees C, although no differences in cell size were observed. The density of MHC Class I and II expression was higher on cells maintained at 39.2 degrees C, as was the percentage of MHC Class II positive cells. In contrast, expression of CD44 and CD11a/18 remained unchanged. Following stimulation with lipopolysaccharide, only cells maintained at 39.2 degrees C produced detectable levels of TNF-alpha. As a final reference criterion, replication of the macrophage tropic African swine fever virus was monitored. At 39.2 degrees C, virus antigen production was less efficient, and virus isolate-related differences in the replication kinetics were observed. Infectious virus production was not different at the two temperatures, implying that virus maturation may have been more efficient at the higher temperature. These results indicate that incubation of cultured cells at the temperature of their donor species has an important influence on their characteristics.