Culture of colony-forming hematopoietic progenitor cells from human peripheral blood

Summary Methods for culturing hematopoietic colonies from human peripheral blood are described in this report. Granulocyte-macrophage progenitor cells (CFU-GM: colony forming units-granulocyte/macrophage) are grown in 35-mm dishes containing 4 ml of 0.35% agarose in Iscove's modified Dulbecco's medium (IMDM) with 20% prescreened fetal bovine serum (FBS). Colony-stimulating activity is provided by leukocyte-conditioned medium, and nonactivated autologous T lymphocytes or recombinant granulocyte- and granulocyte/macrophage-colony stimulating factor or both. This procedure minimize the growth inhibition caused by monocytes. Erythroid progenitor cells (BFUe: burst forming units-erythroid) are cultured in 1 ml of 0.35% agarose in IMDM with 30% FBS and conditioned medium from human bladder carcinoma cell line 5637 or recombinant interleukin 3. Both assays have proved useful in studying the enrichment of peripheral blood hematopoietic cells.