Myeloid differentiation of human blood mononuclear cells in liquid culture

Abstract

Abstract Human blood mononuclear cells were prepared from peripheral blood or single donor plateletpheresis residues by depletion of adherent cells and T lymphocytes. In double-layer soft agar culture, 5 x 10(5) such cells yielded from 33 to 165 colonies. For liquid culture, these cells were suspended in McCoy's 5A medium with 15% fetal bovine serum and 0%, 20%, or 40% conditioned medium (CM) and incubated for up to 14 days at 37 degrees C in a humidified 5% CO2–95% air atmosphere. The number of cells in cultures with CM decreased about 0%-10%, while cell counts from cultures without CM decreased about 45%-65%. In cultures with CM, 5%-20% of the cells were classified as blasts after 3–5 days. After 7- 11 days, blasts and promyelocytes comprised up to 53% of all cells. After 9–11 days, cells with specific granules and maturing nuclei comprised up to 56% of all cells. At 11 days, up to 66% of the cells contained peroxidase-positive granules. Cultures without CM contained no more than 5% blasts and promyelocytes and less than 5% maturing granulocytic cells. 3H-thymidine and Na235SO4 incorporation reached a peak at 3–5 days and at 5–11 days, respectively, in cells from cultures containing CM.