Cellular abnormalities of folate deficiency

To trace the development of folate-deficient abnormalities of morphology and DNA synthesis, Friend erythroleukaemia cells were grown in media containing 10(2), 10(3) and 10(4) ng of [3H]PteGlu1/ml and then transferred to folate-free media. Parameters examined were: intracellular folate levels; growth potential; morphology; dU suppression; and DNA content by flow microfluorimetry. The most sensitive indicators of folate-deficient cell growth were those related to DNA synthesis (dU and flow microfluorimetry). These became abnormal at intracellular folate levels of 0.2-0.5 ng/10(6) cells and markedly so below 0.1 ng/10(6) cells. Morphological criteria were less sensitive. Cells became megaloblastic at intracellular folate levels below 0.06 ng/10(6). The capacity of the cells to replicate in folate-free media was a function of the intracellular folate (ICF): duplications = 4.01 + ln(ICF)/0.67 (r = 0.993, P less than 0.001). These studies demonstrate that regardless of initial intracellular folate levels, cellular stigmata of folate deficiency appear when cellular folate falls below 3 X 10(5) molecules per cell (dU and flow microfluorimetry) and cells lose the capacity for further replication below 7-10 X 10(5) molecules. The intracellular folate level not only predicts early defects, but also determines the replicative capacity.