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Development of an optimized protocol for primary culture of smooth muscle cells from rat thoracic aortas

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Abstract

Primary culture of smooth muscle cells has been widely used as a valuable tool to study the molecular mechanisms underlying atherosclerosis and restenosis. Currently, tissue explants and enzymatic digestion methods are frequently applied to produce smooth muscle cells. Explants method is time consuming, usually taking several weeks. The enzymatic digestion method requires large amounts of proteolytic enzymes to generate enough cells for cardiovascular research. The present study reports an optimized method by combining both techniques to obtain high purity smooth muscle cells. The cultured cells exhibited the characteristic “hills and valleys” growth pattern as observed by phase contrast microscopy and showed α-SM-actin positive staining by indirect immunocytochemistry and immunofluorescence. Purity of the cells is guaranteed by the lack of von Willebrand Factor immunoreactivity. Finally, the cultured cells well proliferate on oxidized-LDL stimulation, suggesting the practical utility of this new method.

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Acknowledgments

This study was supported by research grants from National Natural Science Foundation of China (No: 30472022), and Major Program in Key Field of People’s Government of Guangdong Province (P.R. of China) (No: 2003A30904), and Major Program in Scientific Research of Ministry of Education (P.R. of China) (No: 104146). We are grateful to Dr. Shaorui Chen and Dr. Minfeng Su for expert technical assistance. The authors extend their gratitude to the laboratory personnel for encouragement and support.

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  1. Pingxi Xiao

    Present address: Department of Cardiology, Nanjing Medical University Affiliated Nanjing First Hospital, 210006, Nanjing, People’s Republic of China

Authors and Affiliations

  1. Department of Pharmacology and Toxicology, School of Pharmaceutical Sciences, Sun Yat-Sen University, East of Waihuan Road 132, High Education Mega Center, 510006, Guangzhou, People’s Republic of China

    Suowen Xu, Jiajia Fu, Jianwen Chen, Tian Lan, Kang Le, Lan He, Xiaoyan Shen, Heqing Huang & Peiqing Liu

  2. Department of Cardiology, First Affiliated Hospital of Sun Yat-Sen University, Zhonshan 2 Road, 510080, Guangzhou, People’s Republic of China

    Fei Cheng

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  1. Suowen Xu
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Correspondence to Peiqing Liu.

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Suowen Xu and Jiajia Fu have equally contributed to this work.

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Xu, S., Fu, J., Chen, J. et al. Development of an optimized protocol for primary culture of smooth muscle cells from rat thoracic aortas. Cytotechnology 61, 65–72 (2009). https://doi.org/10.1007/s10616-009-9236-6

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