人非小细胞肺癌血管内皮细胞的分离培养及鉴定

Isolation and culture of human non-small cell lung cancer-derived vascular endothelial cells

目的:建立人非小细胞肺癌血管内皮细胞体外分离培养体系,研究其体外生长特性。方法:利用酶消化法分离出肺癌组织微血管片段,采用植块培养法培养原代内皮细胞,先后以局部消化法和差速黏附法进行纯化;应用光镜、免疫细胞化学及免疫荧光技术对所获得的人肺癌血管内皮细胞进行鉴定。结果:所获人非小细胞肺癌血管内皮细胞呈FⅧ-RAg、CD34阳性;电镜下生长状态良好、可传代培养。结论:本研究摸索并建立了人非小细胞肺癌血管内皮细胞的分离培养方法,所获人非小细胞肺癌血管内皮细胞对肿瘤血管生成及抗肿瘤药物的研究具有重要价值。

Objective： To establish a method for isolating and culturing non-small cell lung cancer-derived vascular endothelial cells （NSCLCDVECs） and to observe their characteristics in vitro. Methods： Enzyme digestion method was used to isolate microvascular fragments from human non-small cell lung cancer tissues,and explant culture method was used for primary culture of the mierovascular endothelial cells, then the cells were purified by local digestion and differential adhesion method. The obtained NSCLCDECs were identified by optical microscope, immunocytochemistry and immunofluorescence methods. Results： The NSCLCDECs were positive for FⅧ-RAg and CD34. Microscopy showed that the cells grew well and could be passaged. Conclusion： We have established a method for isolating and culturing NSCLCKVECs, and the resultant cells possess a great value for research on anti-vascularizaton and anti-tumor agents.