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| A modification of the mouse aortic ring assay for angiogenesis |
| WENG Jia-yi， YAN Jin-chuan, CHEN Yao |
| （Department of Cardiology, Affiliated Hospital of Jiangsu University， Zhenjiang Jiangsu 212001, China） |
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Abstract Objective: To optimize the method of the mouse aortic ring angiogenesis assay. Methods: The thoracic aorta of C57BL/6 mice were separated and all extraneous fat and connective tissues were removed. To remove the vascular adventitia, the aorta was digested with collagenase Ⅱ. The aorta were cut into rings～0.5 mm in width and the rings were embeded in the wells of a 96well plate coated with type Ⅰ collagen. The embedded rings were fed with 100 μL of DMEM culture medium supplemented with 10% FBS or 2.5% FBS and VEGF to attain a final concentration of 50 ng/mL. Inverted microscope was used to observe microvessel outgrowth of the aortic rings. The endothelial cells were assayed by BS1 lectinFITC immunofluorescence. Results: The number of microvessels of the aortic rings in the 96well plate was more than those in 24well plate（t=-6.000，P<0.05）. The number of microvessels of the aortic rings cultured with DMEM+10% fatal bovine serum was more than those cultured with DMEM+2.5% fatal bovine serum or DMEM+2.5% fatal bovine serum+VEGF. Microvessel outgrowth of the aortic rings started at 2 to 4 days and reached the peak at day 6 to 8 using the modified method. Conclusion: A great quantity, purity and high survival rate of the mouse aortic ring model of angiogenesis can be effectively obtained by coating plates with type Ⅰ collagen and enhancing the concentration of FBS.
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Received: 07 January 2016
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2016, Vol. 26 
