EPCs联合胚胎鼠跖骨血管生成实验模型的建立

发布时间：2018-02-03 01:48

本文关键词： EPCs 胚胎鼠跖骨血管生成实验模型　出处：《石河子大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:1.在胚胎鼠跖骨血管生成实验基础上,引入外源性的内皮祖细胞(EPCs),建立一种新型体外血管生成模型。研究者可通过此实验模型直接观察EPCs对胚胎鼠跖骨血管生成的影响,并借助于此实验模型可以更好地筛选或反应不同实验目标对EPCs活性的促进或抑制作用。2.探讨EPCs联合胚胎鼠跖骨血管形成实验模型中加入的EPCs的最适数量、模型的最适共培养时间。方法:1.分离、培养及扩增来源于6-8周龄的C57BL/6小鼠骨髓的内皮祖细胞(EPCs);2.从孕17.5天C57BL/6小鼠中分离提取胚胎鼠,无菌条件下将胚胎鼠跖骨取出;3.分别将数量为5×103、1×104、5×104、1×105的EPCs与胚胎鼠跖骨进行时间分别为12天、14天、16天的联合培养,并设空白对照组;4.CD31对胚胎鼠跖骨周围芽生管样结构进行免疫染色;5.用Image-Pro Plus 6.0对免疫染色阳性结果图像区域进行量化;6.SPSS 17.0分析处理数据并进行统计学分析。结果:1.利用全骨髓贴壁法结合差时贴壁法可有效分离和扩增EPCs细胞。2.EPCs联合胚胎鼠跖骨血管生成实验模型的最适共培养时间为14天。3.EPCs联合胚胎鼠跖骨血管生成实验模型中加入的EPCs相对最适数量为5×104。结论:初步构建了EPCs联合胚胎鼠跖骨血管生成实验模型,并实现对EPCs活性进行定量检测。
[Abstract]:Objective to introduce exogenous endothelial progenitor cells (EPCs) on the basis of angiogenesis of fetal rat metatarsal bone. To establish a new in vitro angiogenesis model, the researchers can directly observe the effects of EPCs on the angiogenesis of the metatarsal bone in embryonic mice. With the help of this experimental model, we can better screen or respond to the effects of different experimental targets on the promotion or inhibition of EPCs activity .2.To explore the EP added in the experimental model of vascularization of the metatarsal bone of EPCs combined with embryo mice. The optimum quantity of Cs. Methods: 1. Isolation, culture and amplification of endothelial progenitor cells from the bone marrow of C57BL / 6 mice aged 6-8 weeks. 2. Embryonic mice were isolated and extracted from C57BL / 6 mice on day 17.5 of gestation, and the metatarsal bones of embryonic mice were removed under aseptic condition. 3. EPCs with a quantity of 5 脳 10 ~ 3 ~ 1 脳 10 ~ 4 ~ 1 脳 10 ~ 4 ~ 1 脳 10 ~ 5 and mouse metatarsal bone were co-cultured for 12 ~ 14 days and 16 days, respectively. A blank control group was set up. 4. CD31 was used for immunocytochemical staining of embryonic mouse perimetatarsal sprouts. 5. Image-Pro Plus 6.0 was used to quantify the image area of immunoreactive results. 6.SPSS 17.0 data processing and statistical analysis. Results:. 1. The best co-culture time was 14 days. 3. The best co-culture time of EPCs cells combined with embryonic mouse metatarsal bone angiogenesis was 14 days. The optimal number of EPCs added in the experimental model of Cs combined with fetal mouse metatarsal bone angiogenesis was 5 脳 10 4. Conclusion:. The experimental model of EPCs combined with fetal rat metatarsal bone angiogenesis was preliminarily constructed. The activity of EPCs was detected quantitatively.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R-332

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