Objective To construct fusion cells with induced pluriopotent stem cells(iPSCs) and cardiomyocytes in vitro,and to investigate biological characteristics of cytogenetic and directed differentiation of fusion cells. Methods Polyethylene glycol(PEG-4000)was used to mediate the cell fusion of iPSCs and primary cardiomyocytes,the distribution of fusion cells chromosome number and alkaline phosphatase staining were detected. Specific genes and proteins of parental cells in fusion cells were identified,and ability to differentiation into cardiomyocytes when fusion cells lose parental cardiomyocytes phenotype. Results The fusion cells showed iPSCs-like morphology,mainly showed tetraploid or near tetraploid karyotype(more than 75% of the fusion cells had chromosomes between 76 and 80). The positive rate of alkaline phosphatase in fusion cells was lower than iPSCs at the same time point. There was no difference expression of stem cell-specific gene Oct-4 and Nanog in the fusion cells and iPSCs,but the expression of cardiomyocytes-specific gene α-MHC and β-MHC in fusion cells was significantly lower than cardiomyocytes. Oct-4 protein was expressed in different generations of fusion cells,whereas with time,cTnT protein negative expression gradually. The fusion cells that lost the parental cardiomyocytes phenotype were able to differentiate into cardiomyocytes,and the differentiation rate was higher than iPSCs at the same time point. Conclusions Fusion cells from iPSCs and primary cardiomyocytes,initially showed bidirectional reprogramming,fully exhibit stem cell phenotype after fifth generation and can differentiate into cardiomyocytes.
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