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| Comparison of different methods for mouse pulmonary microvascular endothelial cells culture |
| ZHOU Lingping, FENG Cheng, TANG Xuebin, XU Honglei. |
Department of Pulmonary and Critical Care Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015
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| Cite this article: |
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ZHOU Lingping,FENG Cheng,TANG Xuebin, et al. Comparison of different methods for mouse pulmonary microvascular endothelial cells culture[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2016, 46(8): 590-593.
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Abstract Objective: To explore an efficient and stable method for the mouse pulmonary microvascular endothelial cells (PMVECs) culture. Methods: Forty-five 1 week old Balb/c mice were randomly divided into three groups. Mouse PMVECs were isolated and cultured respectively by enzyme digestion method, explants method and immunimagnetic beads method. Cells were identified by morphological observation and VIII factor related antigen immunofluorescence staining. The success rate of primary culture and the time required from primary culture to the beginning of the first passage was calculated, the cell purity was determinated, growth curves were drew by MTT method. Results: Short spindle and polygonal microvascular endothelial cells were observed in all of the three methods. Those cells were positive to the VIII factor related antigen. The success rate and cell activity in immunomagnetic beads method group increased significantly compared with the other two groups (P<0.01). As for the time required from primary cultured to the beginning of the first passage, it decreased in immunomagnetic beads method group compared with the explant method group (P<0.05). And the purity increased significantly in immunomagnetic beads method group compared with the enzyme digestion method group (P<0.01). Conclusion: Immunomagnetic beads method is of higher repeatability, rapider separation, and obtains much higher purity and better activity mice PMVECs.
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Received: 08 June 2015
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. [J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2022, 52(8): 0-. |
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