Generation of thalassemia-specific integration-free induced pluripotent stem cells and determination of their differentiation ability / 中国病理生理杂志

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.

Study on 5-carboxyfluorescein N-succinimidyl ester in detecting the degree of cross-linked allogenic tissue / 生物医学工程学杂志

This study was designed to creat a new method for detecting the degree of cross-linked allogenic tissue based on fluorescent technique. The thoracic aorta of New Zealand rabbits were divided randomly into four groups according to the concentration of Glutaraldehyde (GA), which were group A (control group-with no GA), group B (cross-linked with GA of 0.625%), group C (1.25%), and group D (2.5%). Each group was cross-linked with GA and reacted with 5-FAMSE, and then the fluorescence intensity was observed via fluorescence microscopy (analyzed with Image-Pro Plus 6.0, a professional image analysis software). The differences between groups in order of fluorescence intensity were found to be: group A > roup B > group C > group D (P < 0.01). Meanwhile, the tissue proteins extracted from aorta in each group were submitted to conventional polyacrylamide gel electrophoresis (PGE) after being cross-linked with GA; the result from this method was compared with that from the method of 5-FAMSE. In group A, the tissue proteins extracted from the aorta cross-linked with GA were obviously less than those not cross-linked with GA. However, this phenomenom was not clearly seen among the B, C and D groups. Nevertheless, 5-FAMSE can detect the degree of cross-linkage more conveniently and directly.