Generation of thalassemia-specific integration-free induced pluripotent stem cells and determination of their differentiation ability

AIM: To generate thalassemia-specific integration-free induced pluripotent stem cells(iPSC) and to detect their ability of differentiation into hematopoietic precursors.METHODS: The plasmids pEB-C5 and pEB-Tg were transfected into the fibroblast cells from hemoglobin Bart's hydrops fetalis's skin by the method of nuclear transfection to reprogramm the cells into iPSC. The ability of the iPSC to differentiate into 3-germ layer cells was determined. The iPSC were cocultured with mouse OP9 cells to differentiate into hematopoietic precursors and the hematopoietic precursor specific antigens were detected. RESULTS: The integration-free iPSC from hemoglobin Bart's hydrops fetalis's skin fibroblasts were successfully derived, and had the ability to differentiate into 3 germ layers. When cocultured with OP9 cells for 9 d, the positive rate of hematopoietic progenitor cell marker CD34 was 18.7%, and the CD34 and CD45 double positive rate was 12.2%. CONCLUSION: Hemoglobin Bart's hydrops fetalis's skin fibroblasts can be successfully induced into "integration-free" iPSC. This cell line has the ability to differentiate into 3 germ layers, and can be differentiated into hematopoietic precursors when cocultured with OP9 cells.