PHOSPH-HISTONE H3——AN APPROACH TO DETECT THE MITOTIC INDEX OF CARDIOMYOCYTES
- VernacularTitle:磷酸化组蛋白H3——检测心肌细胞有丝分裂指数的可靠指标
- Author:
Jing DI
;
Lingzhi ZHANG
;
Shuling BAI
- Publication Type:Journal Article
- Keywords:
H9c2(2-1) cell;
Mitotic;
Index;
Phospho-histone H3;
?-Sarcomeric actin;
Double immunofluorescence staining;
Rat
- From:
Acta Anatomica Sinica
1954;0(02):-
- CountryChina
- Language:Chinese
-
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(1:100) 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.
