Isolation, identification and culture of porcine heart valve myofibroblasts
10.3969/j.issn.2095-4344.2016.51.013
- VernacularTitle:猪心脏瓣膜成肌纤维细胞的分离、鉴定与培养
- Author:
Fengdan LIU
;
Weilin HU
;
Zhengping CHEN
;
Yongsheng LI
- From:
Chinese Journal of Tissue Engineering Research
2016;20(51):7684-7689
- CountryChina
- Language:Chinese
-
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.
