Optimization of calcification model and primary culture method of mouse aortic vascular smooth muscle cells in vitro

AIM To improve the survival rate and culture efficiency of primary culture of mouse aortic vascular smooth muscle cells (VSMCs) through modified enzyme digestion method, and thus to establish a simple and efficient method of calcification model in vitro.

METHODS The mice aortic medium membrane was peeled off and sheared, and the primary culture of mouse VSMCs was carried out by modified tissue-piece inoculation and modified enzyme digestion method. The purity of cells was determined by immunofluorescence staining and the VSMCs were divided into two groups (classical calcification group and modified calcification group) for in vitro calcification induction. Finally, vonKossa staining was adopted to evaluate the calcification differences between the groups.

RESULTS Compared with the modified tissue-piece inoculation, the modified enzyme-combined digestion could obtain a large number of primary VSMCs in a shorter time and the purity of the identified VSMCs was more than 98%. Compared with traditional calcification methods, the modified method made the calcification model easier and more stable.

CONCLUSION In this study, the survival rate and culture efficiency of primary smooth muscle cells are successfully improved by the modified enzyme digestion method and the preparation of the calcification model in vitro is optimized by improving the formula of the calcification induction solution.