MYELOID DIFFERENTIATION OF HUMAN-BLOOD MONONUCLEAR-CELLS IN LIQUID CULTURE

  • 1 January 1981
    • journal article
    • research article
    • Vol. 57  (3) , 497-504
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 .times. 105 such cells yielded from 33-165 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.degree. 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 Na2 35SO4 incorporation reached a peak at 3-5 days and at 5-11 days, respectively, in cells from cultures containing CM.

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