Isolation, identification and culture of porcine heart valve myofibroblasts / 中国组织工程研究

ABSTRACT

BACKGROUND:Valvular interstitial cel s are the main components of the heart valves. Myofibroblasts, as a kind of valvular interstitial cel s, can express alpha-smooth muscle actin and type I col agen fiber, and hold differentiation potential. These cel s cannot only play a support role in the valve structure, but also play a regulatory role in the process of the valve normal physiological and pathological responses. OBJECTIVE:To obtain a reliable method of separation, primary culture and identification of myofibroblasts laying a foundation for further study on the cardiac valvular calcification. METHODS:Aortic valve myofibroblas extracted from porcine hearts were primary cultured by trypsin and col agenase combined digestive method, common enzyme-digestion method and tissue-culture method, respectively. The myofibroblast activity and morphology were observed using microscope, and myofibroblasts were identified using light microscope and immunocytochemistrial method. RESULTS AND CONCLUSION:Myofibroblasts had a higher activity and purity cultured by trypsin combined with col agenase II digestion method. Aortic valve myofibroblasts were positive for alpha-smooth muscle actin and negative for von Wil ebrand factor under fluorescence microscope, suggesting that myofibroblasts were successful y obtained.