In vitro mass production of human erythroid cells from the blood of normal donors and thalassemic patients

Abstract

We describe a new two-step culture method for mass production in vitro of erythroid cells from either CD34(10cells/mL) or light-density (10cells/mL) cells purified from the blood of normal donors and thalassemic patients. The method includes (i) culture of the cells in the presence of dexamethasone and estradiol (10M each) and (ii) the growth factors SCF (50 ng/mL), IL-3 (1 ng/mL), and EPO (1 U/mL). In their proliferative phase, these cultures generated about 1-2×10erythroblasts for each milliliter of blood collected from normal donors or thalassemic patients. They were composed mostly (90%) of CD45/glycophorin (GPA)/CD71cells at day 7, 50-60% of which became CD45/GPA+/CD71by days 15-20. However, when cells from days 7 to 12 of the proliferative phase were transferred in differentiation medium containing EPO and insulin, they progressed to mature erythroblasts (>90% benzidineand CD45/GPA/CD71) in 4 days. Because of the high number of erythroid cells that are generated from modest volumes of blood, this method will prove useful in donor-specific studies of erythroid differentiation.