PURIFICATION OF HUMAN PERIPHERAL-BLOOD COLONY-FORMING CELLS (CFU-C)

  • 1 January 1980
    • journal article
    • research article
    • Vol. 8  (9) , 1086-1093
Abstract
A 10-fold enrichment of colony-forming cells (CFUC) from single donor plateletpheresis residues and from 70-120 ml of peripheral blood of normal donors was achieved by sequential sedimentation on Ficoll-diatrizoate, depletion of cells adherent to plastic and depletion of cells rosetting with sheep red blood cells (T lymphocytes). Cultures of 5 .times. 105 unfractionated mononuclear cells yielded 9 .+-. 3 colonies and mononuclear cells depleted of adherent cells and T lymphocytes yielded 53 .+-. 6 colonies. The mononuclear cell fraction depleted of adherent cells and T lymphocytes was further enriched for CFUC by isopycnic sedimentation of Percoll gradients. Cells recovered in the 1.063-1.065 g/cm3 density layer of the gradient formed 146 .+-. 9 colonies in culture. The mononuclear cells depleted of adherent cells and T lymphocytes were also enriched for CFU by depletion of Fc-receptor positive cells using an immune sheep red blood cell rosette sedimentation technique. Cultures of the Fc-receptor depleted fractions yielded 107 .+-. 12 colonies, while the Fc-receptor enriched fraction yielded only 2 .+-. 1 colonies. CFUC appear to lack surface membrane receptors for sheep erythrocytes and the Fc portion of Ig as well as the ability to adhere to plastic.