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An improvement in primary culture of rat vascular smooth muscle cells and its identification |
1.Department of Cardiology, Shaoxing People’s Hospital, Shaoxing Hospital of Zhejiang University, Shaoxing, 312000; 2.The First Clinical Medical College, Wenzhou Medical University, Wenzhou, 325035; 3.Zhejiang Chinese Medical University, Hangzhou, 310000 |
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Cite this article: |
MENG Liping1,2,JIANG Chengjian1, et al. An improvement in primary culture of rat vascular smooth muscle cells and its identification[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2015, 45(8): 593-.
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Abstract Objective: To explore a new effective way on primary culture of rat vascular smooth muscle cells (VSMCs), which could shorten the cultivate time and enhance the VSMCs’ purity. Methods: The primary culture of rat VSMCs was conducted by a new way reformed from the traditional modified tissue explant technique, in which the medial layer was isolated and 10 μg/L PDGF-B was added to the high glucose DMEM. The VSMCs were identified by cell morphology and immunocytochemistry. Results: About six day later, VSMCs could be found around some tissue blocks, and passage of the VSMCs can be done about 12 days later. Under the phase contrast microscope, the VSMCs looks like spindle and the specific “hill and valley” phenomenon could be seen. Immunocytochemistry showed that there was a large number of smooth muscle actin (SMA) and the VSMCs’ purity was much higher than the cells got by the old way. Conclusion: Adding PDGF-B could shorten the cultivate time of primary VSMCs and isolation of the medial layer could improve the VSMCs’purity.
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Received: 15 December 2014
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