A S uccessful Generation of Erythroid Precursors from HbE / Beta Thalassemia Patients using Peripheral Blood Mononuclear Cells

Thalassemia is a red blood cells disorder. In diseases related to ineffective erythropoiesis such as thalassemia, it is important to grow erythroid precursor cell (EPC) for the used in research. It would be a breakthrough if minimal amount of blood volume is used to grow EPC from peripheral blood mononuclear cells especially in patients with chronic anemia and transfusion dependent. The purpose of this study was to generate erythroid precursors from peripheral blood mononuclear cells (PMNCs). 2.5ml of whole blood were collected from both normal and HbE/beta thalassemia patients. 1-2 million PMNCs were plated in 2ml of complete erythroid expansion medium. The medium was changed every 2 days. Flow cytometry was performed on day 7 and day 14 to evaluate the erythroid differentiation. We have succeeded in obtaining a good yield of 98% and 99.5% erythroid precursors from normal and thalassemic blood, respectively. Majority EPC grown from thalassemia patients exhibited immature erythroid precursor and double positive for CD 71 and GYPA. This difference in differentiation capacity between normal and thalassemic sample is probably related to the underlying defective erythropoiesis in thalassemic patients. More study of erythropoiesis can be performed using EPC generated from minimal amount of peripheral blood.