Effects of EPO and EST on erythroid maturation

Transcriptional profiles of human CD34+ cells cultured in EPO and EST conditions. After obtaining informed consent, human CD34+ cells were isolated in high purity from the peripheral blood of normal human volunteers. The cells were cultured at a concentration of 104 - 105 cells/mL in medium supplemented with 4 U/mL Erythropoietin (EPO) versus 4 U/mL Erythropoietin + 50 ng/mL Stem Cell Factor + 1.25 ng/mL Transforming Growth Factor Beta (EST) as described previously. Cells were sampled on culture Day 7, culture Day 14, or both culture days and enumerated using an electronic cell counter (Coulter, Hialeah, FL). Adult and fetal hemoglobin was analysed by HPLC. RNA was extracted using QIAshredder and Rneasy Minikit.