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| The influence of two vascular smooth muscle cell primary culture methods on cellular contractile phenotype |
| ZHOU Changzuan1, GUO Hangyuan1,2, MENG Liping1,2, JI Zheng2 |
1.The First Clinical Medical College, Wenzhou Medical University, Wenzhou, 325035; 2.Department of Cardiology, Shaoxing People’s Hospital, Shaoxing Hospital of Zhejiang University, Shaoxing, 312000
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| Cite this article: |
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ZHOU Changzuan,GUO Hangyuan,MENG Liping, et al. The influence of two vascular smooth muscle cell primary culture methods on cellular contractile phenotype[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2016, 46(12): 859-864.
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Abstract Objective: To explore the difference between collagenase digestion method and explant-culture method to obtain vascular smooth muscle cells (VSMCs) primary culture on maintaining contractile phenotype. Methods: The 1st, 2nd, 4th, 8th, 12th generation of primary VSMCs were obtained via type II collagenase digestion and explant-culture method. VSMCs contractile phenotype protein markers, calopnin and smooth muscle actin-α (SMA-α), as well assecreted protein osteopontin (OPN), were detected by western blot between different generations. The migration and proliferation ability of VSMCs from two methods were detected by wound-healing assay and MTT assay, and the phenotype protein markers expression by immunofluorescence and western blot. Results: We found VSMCs obtained from enzyme digestion initially exhibited stronger SMA-α and calponin expression, lower OPN expression and suppressed cellular proliferation and migration compared with that from explant-culture method. However, we observed a positive correlation between cell generation and VSMCs dedifferentiation degree, and a stronger dedifferentiation trend in enzyme digestion cells after 8th generation. Conclusion: Type II collagenase digestion method shows advantages in acquiring primary cells rapidly and maintaining VSMCs contractile phenotype within 4th generation compared with explant-culture method.
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Received: 21 January 2016
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