Effects of five recombinant hematopoietic growth factors on enriched human erythroid progenitors in serum-replaced cultures

✍ Scribed by Maria Teresa Mitjavila; Masaki Natazawa; Pascale Brignaschi; Najet Debili; Janine Breton-Gorius; William Vainchenker

• Publisher: John Wiley and Sons
• Year: 1989
• Tongue: English
• Weight: 786 KB
• Volume: 138
• Category: Article
• ISSN: 0021-9541
• DOI: 10.1002/jcp.1041380324

✦ Synopsis

Erythroid progenitors from normal human marrow were purified by a two-step immune panning method permitting both the enrichment of erythroid progenitors (plating efficency up to 10%) and the separation of CFU-E from BFU-E. The purified erythroid progenitors wcre grown in serum-replaced conditions; in some cxperiments at an average of one cell pcr well. Human recombinant granulocytemacrophage colony stirnulaling factor (GM-CSF), interleukin 3 (IL3), erythroid potentiating activity (EPA), and hurnan erythropoietin (Epo) either recombinant or homogenous native were tested for their effect on CFU-E growth. Epo was an absolute requirement for CFU-E growth and was sufficient to obtain colony formation at the iinicellular level wl-iereas GM-CSF and IL3 did not further increase the plating cfficiency. €PA potentialed the effect of Epo on this progenitor only in experiments pcrformed at unicellulx Icvel. Human recombinant CM-CSF, IL3, Interleukin . I cy (ILlw), and Epo were subsequcntly tested for their ability to proinole BFU-E growth. GM-CSF and IL3 supported the growth of erythroid bursts in the presence of Epo, even at the unicellular level. However, IL3 promoted a higher number of bursts than GM-CSF under all conditions te3ted. These ~w o growth factors have no or very small additive ttffccts when tested in combination. I L l a added to Epo alone had no effect on the growth of BFU-E whereas it potentiated the combined action of IL3 and GM-CSF on the primitive BFU-E. In conclusion, this study confirms at the unicellular level and under serum-free conditions that erythroid progenitors are regulated by multipotential growth factors in early phases ol erythropoiesis and become sensitive only lo Epo in later phases of differentiation.