小鼠主动脉环血管新生实验模型的优化

摘要
图/表
参考文献
相关文章 (2)

全文: PDF (3662 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要 目的: 建立优化体外培养小鼠主动脉环血管新生实验模型的条件和方法。方法: 分离C57BL/6小鼠的胸主动脉，剔除主动脉周围的脂肪及结缔组织，使用Ⅱ型胶原酶消化动脉，剥离血管外膜，将血管剪成0.5 mm长的动脉环，置于预先包被有鼠尾Ⅰ型胶原的96孔板中培养，给予DMEM+2.5%胎牛血清，DMEM+10%胎牛血清或DMEM+2.5%胎牛血清+血管内皮生长因子(VEGF，终浓度为50 ng/mL)100 μL/孔，覆盖胶原表面。倒置显微镜下观察动脉环新生微血管。免疫荧光染色鉴定内皮细胞。结果：鼠尾Ⅰ型胶原包被的96孔板（DMEM+2.5%胎牛血清）内新生微血管数量明显多于24孔板（t=－6.000，P<0.05）。DMEM+10%胎牛血清组新生微血管数量明显高于DMEM+2.5%胎牛血清组和DMEM+2.5%胎牛血清+VEGF组。DMEM+10%胎牛血清组培养2~4 d后小鼠主动脉环开始出芽， 6~8 d后动脉环新生微血管分支、数量及长度达到高峰。结论：优化后的方法可以得到生长状态良好，出芽率高，纯度高的离体小鼠动脉环血管新生实验模型。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	翁嘉懿
	严金川
	陈瑶

关键词 ：主动脉环, 血管新生, 组织培养

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 96well 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 lectinFITC immunofluorescence. Results: The number of microvessels of the aortic rings in the 96well plate was more than those in 24well 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.

收稿日期: 2016-01-07

基金资助:

国家自然科学基金资助项目（81170279, 81370409）；江苏省科教兴卫工程项目（LJ201116）；江苏省六大人才高峰项目（WS074）；江苏省333高层次人才项目（BRA2014162）

通讯作者: 严金川（通讯作者），博士，博士生导师，E-mail: yanjinchuan@hotmail.com

作者简介: 翁嘉懿（1991—），女，硕士研究生

引用本文:

翁嘉懿, 严金川, 陈瑶. 小鼠主动脉环血管新生实验模型的优化[J]. 江苏大学学报:医学版, 2016, 26(03): 227-. WENG Jia-Yi, Yan-Jin-Chuan, Chen-Yao. A modification of the mouse aortic ring assay for angiogenesis. Journal of Jiangsu University(Medicine Edition), 2016, 26(03): 227-.

链接本文:

http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2016/V26/I03/227