Establishment of the thalassemia patient-specific induced pluripotent stem cells from peripheral and cord blood ;cells and its hematopoietic differentiation
ZENG Min-hui
JIANG Man-bo
WEN Yan-fei
PENG Cheng
ZHANG bin
CAI Liu-hong
Abstract:Objective To investigate the construction and hematopoietic differentiation of induced pluripotent stem cells ( iPSCs) from thalassemia patients .Methods Cord blood from artificial labored fetus with HbH disease (α-thalassemia) and peripheral blood from a case of heterozygote of β-thalassemia (genotype IVS-II-654/CD17) were obtained.The iPSCs were re-programed from mononuclear cells separated from obtained cord blood and peripheral blood . Hematopoietic stem cell differentiation was induced by OP 9 cell co-culture.The pluripotent markers of iPSCs were detec-ted by immunofluorescence and alkaline phosphatase staining , while embryoid bodies ( EBs) and teratoma forming experi-ments were used to detect the pluripotency .Results Pluripotent biomarkers , including Oct4, SSEA4, tra-1-60 and tra-1-81, and alkaline phosphatase , were positive on iPSCs.EBs were formed by suspended culture .Three germ layer cells were found in the teratoma test .The results of flow cytometry showed that CD 34+cells was 4.46%and 8.71%from cord blood and peripheral sources , respectively, after co-cultured with OP9 cells.Conclusion Cord blood and periph-eral blood MNCs of patients with αorβthalassemia, can be reprogrammed into iPSCs , and differentiated into hematopoi-etic stem cells.
Keywords:thalassemiainduced pluripotent stem cellshematopoietic differentiation
Publication Date:2015-01-01
Online Publishing Date:2025-08-15(First online date of this platform, not the publication date of the document)
Pages:5( 1633-1637 )
