Enhancement of cell production in long-term bone marrow culture

In an effort to increase the long-term production of hematopoietic cells in vitro, Origen hybridoma cloning factor (HCF) was added at the initiation of Dexter type cultures, in which whole bone marrow (BM) was seeded into tissue culture flasks and formed an adherent stromal layer that supported the proliferation and differentiation of primitive cells. After about six weeks, all the cultures were fully established, and continuous production of nonadherent cells was maintained for at least 27 weeks. In the groups with 20% HCF, there was a significant (three- to fourfold) increase in the steady-state cell production of 106 +/- 17 x 10(4) cells/ml compared to 26 +/- 10 x 10(4) in controls. In some cases the ability of HCF to increase productivity was limited by the nutrients and metabolic products in the culture medium. Cell number varied inversely with glucose and pH. HCF increased the concentration and absolute number of myeloid progenitors (granulocyte-macrophage colony forming units and spleen colony forming units) in the nonadherent layer and shifted the differentiation of granulocyte-macrophage colony forming units toward the production of cells of the monocyte/macrophage lineage. Spleen colonies produced from 10(5) cells from cultures with HCF were more numerous (8 +/- 2 versus 4 +/- 2) and larger than those from control cultures (2.6 versus 0.2 mg/colony), but they contained the usual cell lineages (erythrocytic, granulocytic and megakaryocytic).