Fractionation of subsets of BFU-E from normal human bone marrow: responsiveness to erythropoietin, human placental-conditioned medium, or granulocyte-macrophage colony-stimulating factor.
Normal human bone marrow mononuclear cells were fractionated by differential adherence, immunomagnetic separation, and fluorescence-activated cell sorting (FACS). The resultant fractionated cells were cultured in semisolid medium to monitor the presence of BFU-E, Mix-CFC, and nonerythroid CFC. Two populations of cells were recovered on the basis of binding by the monoclonal antibody (MoAb) RFB-1. One of these populations contained BFU-E that were stimulated only by erythropoietin (Epo), whereas the second population contained BFU-E responsive to Epo, Epo and recombinant human granulocyte-macrophage colony-stimulating factor (rHGM-CSF), or Epo and human placental-conditioned medium (HPCM). Prior enrichment of clonogenic cells by removal of adherent and Leu-M3+ve, Leu-4+ve, Leu-7+ve, B1+ve, WEMG1+ve, and Glycophorin A+ve cells, followed by FACS fractionation on the basis of RFB-1 binding, consistently resulted in recoveries of BFU-E, Mix-CFC, and nonerythroid CFC of greater than 100% (up to 800%). These procedures also resulted in enrichment of up to 200-fold and frequencies of 1:6 for BFU-E, 1:5 for CFC, and 1:130 for Mix-CFC.
Kannourakis
G
G
Walter and Eliza Hall Institute of Medical Research, Victoria, Australia.
Johnson
G R
GR
D000911:Antibodies, Monoclonal / Q000276:immunology; D001854:Bone Marrow Cells; D002469:Cell Separation / Q000379:methods*; D003115:Colony-Stimulating Factors / Q000494:pharmacology*; D003470:Culture Media; D004912:Erythrocytes / Q000145:classification* / Q000187:drug effects / Q000276:immunology; D004921:Erythropoietin / Q000494:pharmacology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006133:Growth Substances / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000145:classification* / Q000187:drug effects / Q000276:immunology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000145:classification; D008027:Light; D010920:Placenta / Q000378:metabolism*; D012542:Scattering, Radiation
D000911:Antibodies, Monoclonal; D003115:Colony-Stimulating Factors; D003470:Culture Media; D006133:Growth Substances; D004921:Erythropoietin; D016178:Granulocyte-Macrophage Colony-Stimulating Factor