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Effects of erythropoietin and recombinant cytokines on colony formation and self-renewal by erythroid burst-forming units and granulocyte-macrophage progenitors from mobilized peripheral blood progenitor cells.

The effects of erythropoietin and recombinant cytokines (G-CSF, SCF, IL-3 and GM-CSF) on colony formation and self-renewal by erythroid burst-forming units (BFU-E) and granulocyte-macrophage progenitors (CFU-GM) from mobilized peripheral blood progenitor cells (PBPCs) of the normal donors and patients were investigated. To better understand how combinations of the cytokines may be appropriate for stem cell manipulation, a model involving replating of individual BFU-E and CFU-GM colonies has been used to study the effects of cytokines on colony formation and self-renewal. Based on granulocyte colony-stimulating factor (G-CSF) and erythropoietin(EPO) alone serve as a baseline with which to compare the effects of the combinations with stem cell factor (SCF), interleukin-3 (IL-3) or granulocyte-macrophage colony-stimulating factor (GM-CSF). The results revealed that: 1. BFU-E derived from PBPCs produced a significantly great numbers of subcolonies in EPO plus IL-3 and EPO+SCF+IL-3, and a difference in EPO+SCF compared with EPO alone. 2. Compared patients to normal donors the self-renewal ability of BFU-E was not influenced after EPO plus SCF or IL-3. 3. There was a significantly increased frequency of CFU-GM in the presence of SCF, IL-3, and GM-CSF used in conjunction with G-CSF. Comparing the frequencies of CFU-GM in patients and donors, patients' PBPCs were more sensitive to G-CSF+SCF and GMix (G-CSF+SCF+IL-3+GM-CSF), especially to G-CSF alone than donors. 4. There was a significant increase in AUC (area under the curve of subcolony distribution) in the presence of G-CSF combined with other cytokines. GMix was identified as the optimal combination of cytokines for both the expansion of CFU-GM as well as the expansion of clonogenic progenitor cells. 5. Donors have a high AUC than patients, especially there was a significant increase AUC in G-CSF alone (P = 0.0067).