Fig. 1

Experimental desing of erythroid differentiation of CD34⁺ cells. CD34⁺ cells were isolated from bone marrow (BM) or buffy coat preparation from peripheral blood (PB) and plated in methylcellulose medium supplied with erythropoietin (EPO), stem cell factor (SCF), and interleukin 3 (IL3), and cultured for 6 days (cell expansion period). The resulting were transferred to liquid culture composed by alpha MEM supplied with fetal bovine serum (FBS), 2-mercaptoethanol, EPO, holotransferrin, and bovine serum albumin (BSA). Cells were culture for an additional 6 days (cell differentiation period). Cells were submitted to collection of RNA on days 6, 8 and 12 for quantitative PCR experiments, and to immunophenotyping on days 6 and 12. The dot plots illustrate flow cytometry analyzes for glycophorin A (GPA) and CD71 (transferrin receptor) staining of an erythroid differentiation experiment at day 6 and 12 of a healthy donor. Figure was produced using Servier Medical Art (http://www.servier.com)