Abstract: AIM: To find the method of dissociating and culturing the cardiac stem cells (CSC), purify CSC by cell surface markers of stem cell with c-kit, and induce CSC differentiation into cardiomyocytes by 5-azacitidine (5-aza) in vitro.METHODS: The heart cells were dissociated and extracted by zymo-timing digestion and cultured in vitro, the c-kit masculine cells as CSC markers were purified by immunomagnetic beads method.The expression such as c-kit, lin and CD₃₄ as CSC markers were detected after purification by flow cytometry.The cardiac specific troponin T (cTnT), connective-43 (Cx-43) and the specificity desmin of cardiomyocytes were detected by the method of immunocytochemical stain.The differences of the results in the different conditions were compared and analyzed combined with the morphology changes.RESULTS: The cells, which were dissociated and digested from C57BL mouse cardiac muscles, were cultured in vitro. There was a kind of small, round and high-photonasty cells.The speed of cell multiplication after serial subcultivation was faster than the primary cell.These cells were similar to the stem cell from their cell multiplication and the morphological feature.Highly purified ckit + CSC were separated by immunomagnetic beads method, but the CSC do not express CD₃₄ and lin generally.Under the different inducing conditions, these CSC could differentiate into pulsatory cells, and express the specific protein of cardiomyocytes.CONCLUSION: The CSC can be obtained from cardiac muscular tissues of C57BL mice, the highly purified CSC of c-kit +-CD₃₄^--lin^- can be separated by using the magnetic activated cell sorting (MACS) system. After induced by 5-aza or changed the microenvironment of CSC growth, the CSC can differentiate into cardiomyocytes and form " cardiomyocytes cluster" construction.

Key words: cardiacstemcell, induction, differentiation, cadiocyte, 5-azacitidine