Growth of erythroid cells from thawed unseparated cord blood in vitro without exogenous erythropoietin.

Introduction: Previous erythroid cell cultures have depended on added serum or erythropoietin. In this paper, the growth of erythroid cells from thawed unseparated cord blood units in vitro without serum or exogenous erythropoietin is reported.

Methods: Thawed volume-reduced cord blood was cultured in conditions designed to support the megakaryocytic lineage, with thrombopoietin and interleukins 3 and 6. Erythroid cells were detected with glycophorin A (GlyA), CD71, and benzidine (flow cytometry and immunocytochemistry).

Results: Nucleated and anucleated GlyA-positive, as well as benzidine-positive cells were observed from day 9. In flow cytometry, at days 0 and 9, 5.9% and 14% of all events were GlyA+, and 14% and 53% were CD71+, respectively. At days 0 and 9, 4.5% and 12% of the events were double-positive for GlyA and CD71, respectively. By day 14, the percentages of GlyA+, CD71+ and double-positive events had started to decrease (9.7%, 35%, and 5.3%, respectively).

Conclusions: Erythroid cells were generated from thawed unseparated cord blood units without exogenous erythropoietin. Thawed cord blood possesses the potential for erythroid growth in vitro in a culture medium designed for other cell types.