Comparative ound analysis effects of separated cardiomyocytes by collagenase Ⅰ and Ⅱ

doi:10.3969/j.issn.0529-1356.2013.06.010

Abstract

Abstract:

Objective To explore the role of collagenase Ⅱ in the separation of adult rat cardiomyocytes. Methods Collagenase Ⅰ and collagenase Ⅱ were used to digest cardiomyocytes and their effect of separating cardiomyocytes was compared. Immunohistochemistry, immunofluorecence and Western blotting were used to detect existence of collagen Ⅱ in adult rat myocardium. Collagen Ⅱ of xiphoid and collagen Ⅲ of myocardium were taken as positive control. SDS-PAGE(polyacrylamide gel electrophoresis) method was used to detect the composition of collagenase Ⅱ. Results After digesting cardiomyocyte with collagenase Ⅰ or collagenase Ⅱ, the number of the obtained myocardial cells were (5.66±0.83)×106 and (5.61±1.01) ×106, the survival rates of cardiomyocyte were（74.17±6.27）％and（75.50±9.07）%, respectively, and no significant difference was found. Most separated cardiocytes did not beat while only very few of them had automaticity systole; Collagen Ⅱ showed negative expression in myocardium of adult rats but positive expression in cartilaginous tissue of xiphoid; Collagen Ⅲ also showed positive expression in myocardium. Western blotting did not show the expression of collagen Ⅱ in myocardium. SDS-PAGE results showed collagenase Ⅱ was a mixture of five kinds of protein. Conclusion The collagen Ⅱ did not exist in myocardium, thus collagenase Ⅱ should not be used to separate adult rat cardiomyocyte.

Key words: Collagenase Ⅱ, Collagen Ⅱ, Myocardium, Immunohistochemistry, Immunofluorecence, Rat

WANG Hui-ling GUO Kang GUO Zhi-kun*. Comparative ound analysis effects of separated cardiomyocytes by collagenase Ⅰ and Ⅱ[J]. AAS, 2013, 44(6 ): 784-788.

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