PHOSPH-HISTONE H3——AN APPROACH TO DETECT THE MITOTIC INDEX OF CARDIOMYOCYTES / 解剖学报

ABSTRACT

Objective Applying a reliable precise method to assess the mitotic index of cardiomyocytes,to disclose of disclosure the mechanism implicated in cardiomyocytes proliferation. Methods H9c2(2-1) cardiomyocytes were originally developed from rat BD1X heart(ATCC).These cells were cultured on coverslips.Double immunofluorescence staining with monoclonal antibodies(1100) against phospho-histone H3 and ?-sarcomeric actin was performed on the cultured cells.Anti-mouse IgG FITC was used as the secondary antibody for the H3P antibody,and anti-mouse IgM Cy3 was used as the secondary antibody for the ?-sarcomeric actin.DNA was visualized with Hochest 33342.All photographs were taken with an Olympus fluorescence microscope. Results The cytoplasm of cardiomyocytes appeared red,the mitotic chromosomes green with distinct shape,and Hochest 33342-stained nuclei blue.Conclusion Our method is the reliable and exact means to observe and assess cardiomyocytes mitosis.