The activities of 5-methyltetrahydrofolate (5-CH3-THF)-related enzymes [5-CH3-THF homocysteine methyltransferase and 5,10-methylenetetrahydrofolate (5,10-CH2-THF) reductase] and DNA polymerase alpha were measured in normal and malignant hematopoietic cells. The 5-CH3-THF homocysteine methyltransferase activity was significantly correlated with 5,10-CH2-THF reductase activity, indicating that the hematopoietic cells with active biosynthesis of tetrahydrofolate from 5-CH3-THF also actively synthesize 5-CH3-THF from 5,10-CH2-THF. The activities of 5-CH3-THF-related enzymes had a tendency to be high in lymphoid cells and low in myeloid cells, and were not correlated with the percentage of blasts and immature cells in the samples examined. Fairly good correlations were observed among these three enzymes in non-malignant bone marrow cells. However, the activities of two of the enzymes correlated only weakly overall with DNA polymerase alpha activity in normal and malignant hematopoietic cells. Generally speaking, DNA polymerase alpha activity correlated well with the percentage of blasts and immature cells in the samples examined.