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Understanding the major pathways in the process of cardiogenesis is crucial for controlling fate of human embryonic stem cells(hESCs and optimizing cardiac differentiation strategy which could provide sufficient cells for cellular therapy. To assess the role of APJ in human cardiac differentiation.
After monolayer-based cardiac cell culture,hESCs were changed to cardiac differentiation medium which were chemically defined and small molecule-based .In the process of cardiac differentiation,the expression of Brachyury T, mesp1, Nkx2.5 and APJ protein and mRNA were detected using confocal microscope and real-time PCR respectively.
Beating cardiac cells appeared at d7 and it's number didn't increase till d16. After purification of cardiomyocytes, protein of a-actinin and cTNNT2 were positively expressed and sarcomere and intercalated discs were observed obviously. Cyto-immunofluorescence staining showed that APJ was co-expression with stage-specific markers of Brachyury T, mesp1, Nkx2.5 in d2, d3, d7。The mRNA expression of APJ was persistent in all three stages, highest in mesoderm and then decrease gradually.
APJ was co-expression with stage-specific markers of Brachyury T, mesp1, Nkx2.5 and APJ was persistently expressed in the process of human myocardial development. APJ may be a new marker of cardiac progenitor cells.