Generation of thalassemia-specific integration-free induced pluripotent stem cells and determination of their differentiation ability
10.3969/j.issn.1000-4718.2015.02.010
- VernacularTitle:地中海贫血“integration-free”诱导多能干细胞的建立及造血分化的研究
- Author:
Manbo JIANG
;
Minhui ZENG
;
Jun ZAHNG
;
Yanfei WEN
;
Bin ZHANG
;
Liuhong CAI
- Publication Type:Journal Article
- Keywords:
Thalassemia;
Induced pluripotent stem cells;
Integration-free;
Hematopoietic differentiation
- From:
Chinese Journal of Pathophysiology
2015;(2):245-249
- CountryChina
- Language:Chinese
-
Abstract:
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 OP 9 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 CD 34 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 “in-tegration-free” iPSC.This cell line has the ability to differentiate into 3 germ layers , and can be differentiated into hemato-poietic precursors when cocultured with OP 9 cells.
