Mice that are homozygous for the autosomal recessive mutation osteopetrosis (op) suffer from a general skeletal sclerosis, and the numbers of macrophages in various tissues are significantly decreased. We report that microglia in op/op mice are not affected by the mutation. They have normal morphology and are present in the CNS in normal frequency. In cultures, disaggregated cells of neopallia can form microglia, but such cells from neopallia of op/op mice form microglia only when colony-stimulating factor 1 (CSF-1) is added to the culture medium. The addition of granulocyte/macrophage (GM)-CSF or interleukin (IL)-3 to the culture medium does not stimulate production of microglia. Microglia that form in op/op neopallial cell cultures, in the presence of CSF-1, are capable of Fc-receptor-mediated phagocytosis. Based on our experiments, it seems that microglia are CSF-1 dependent but in op/op mice (in which CSF-1 is absent) microglia may use other locally produced factors.