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目的探索和建立一种高效的人脐动脉内皮细胞分离和培养方法。方法用PBS灌注清洗脐动脉后,以0.1 g/L1型胶原酶消化脐动脉内膜,收集、离心消化液,重悬细胞在特定培养基中培养。观察其形态特点,同时用CD31免疫荧光染色和内皮细胞管状结构形成实验对所得细胞进行鉴定。结果倒置相差显微镜下观察所获得的细胞为单层生长,呈现铺路石样形态,并且大量表达内皮细胞特异性膜蛋白CD31,同时能够形成明显的管状结构。结论本研究建立了操作简单、快速、高效分离人脐动脉内皮细胞的方法,所分离得到的内皮细胞纯度高、成活率高。
Objective To explore and establish an efficient method of human umbilical artery endothelial cell isolation and culture. Methods After umbilical artery was infused with PBS, umbilical artery intima was digested with 0.1 g / L collagenase. The digestive juice was collected, centrifuged and resuspended in culture medium. Morphological characteristics were observed. The cells were identified by CD31 immunofluorescence staining and endothelial cell tubular structure formation assay. Results The cells obtained under inverted phase contrast microscope showed monolayer growth, showing paving stone-like morphology and abundant expression of endothelial cell-specific membrane protein CD31, with obvious tubular structure. Conclusion This study established a simple, rapid and efficient method of human umbilical artery endothelial cells isolated, the isolated endothelial cells with high purity and high survival rate.