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19 August 2017 : Laboratory Research  

Primary Culture of Rat Aortic Vascular Smooth Muscle Cells: A New Method

Jufang Chi1ABCDEF, Liping Meng1AB, Sunlei Pan2AB, Hui Lin2BCDE, Xiaoya Zhai1DE, Longbin Liu1B, Changzuan Zhou2D, Chengjian Jiang1BC, Hangyuan Guo12ABCG*

DOI: 10.12659/MSM.902816

Med Sci Monit 2017; 23:4014-4020

Abstract

BACKGROUND: Developing a simple and efficient method of obtaining primary cultured VSMCs is necessary for basic cardiovascular research.

MATERIAL AND METHODS: The procedure of our new method mainly includes 6 steps: isolation of the aortic artery, removal of the fat tissue around the artery, separation of the media, cutting the media into small tissue blocks, transferring the tissue blocks to cell culture plates, and incubation until the cells reach confluence. The cells were identified as VSMCs by morphology and immunofluorescence. Then, VSMCs obtained by this new tissue explants method, the traditional tissue explants method, the enzyme digestion method, and A7r5 cell line were divided into 4 groups. The purity of cells was test by multiple fluorescent staining. Western blotting was used to investigate the phenotype of VSMCs obtained by different methods.

RESULTS: Cells began to grow out at about 8 days and became relatively confluent within 16 days. Compared with VSMCs from the traditional tissue explants method and enzyme digestion method or A7r5 cell line, VSMCs obtained by our method showed higher purity and manifested a more “contractile” phenotype characteristic.

CONCLUSIONS: We have conquered the disadvantages in the previous primary culture methods and established a simple and reliable way to isolate and culture rat aortic VSMCs with high purity and stability.

Keywords: Muscle, Smooth, Vascular, Phenotype, primary cell culture

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750