Sequential study on spontaneous colony formation by bone marrow cells during butylnitrosourea-induced leukemogenesis in the rat.

The spontaneous colony (SC)-forming activity of bone marrow cells of rats during butylnitrosourea (BNU) treatment was studied sequentially in an attempt to analyze stages of leukemogenesis. Aspirated bone marrow cells from female Sprague-Dawley (SD) rats that had been given continuous access to drinking water containing 400 ppm BNU were examined at intervals of 3-5 weeks for colony formation of granulomonocytic cells with or without supplemental colony-stimulating factor (CSF). Granulocytic leukemia was first observed at week 12, and the cumulative incidence reached 80% by week 30. SCs were obtained in 56% of rats in the early stage (3 weeks) and in up to 59% of rats in the late stages (20-25 weeks). However, in the middle stages colony formation was rare, even with the addition of CSF. When adherent cells were removed from the bone marrow cells, the SC-forming activity in the early stage was almost entirely lost, whereas much of that in the late stage remained. It is possible that in the former case, overproduction of endogenous CSF by adherent cells under the influence of BNU treatment could be involved. In contrast, late stage SC formation may be associated with the generation of altered cells, including leukemic or preleukemic elements, which have increased capacity for autonomous growth. The loss of SC-forming activity in the middle stage appeared to be attributable to an extreme reduction in endogenous CSF due to marked devastation of the bone marrow. Technical improvement in adjusting more precisely the level of CSF in the culture medium is required to enable further analysis of leukemogenesis, focused on the colony-forming activity of target cells.