人非小细胞肺癌微血管内皮细胞原代培养方法改进  

A modified method for in vitro isolation and cultivation of Non-small lung cancer-microvascular endothelial cells in Human

作　　者：张峤[1] 郝少欢[2] 吴梅[1] 帕提古丽.阿尔西丁 ZHANG Qiao;HAO Shaohuan;WU Mei;Patiguli Aerxiding(Lung cancer No.1 Ward,The Tumor Affiliated Hospital,Xinjiang Medical University,Urumqi 830011,Chian;Department of Oncology,The Fist People′s Hospital of Kashgar,Kashgar Xinjiang 844000,China)

机构地区：[1]新疆医科大学附属肿瘤医院肺癌一病区,乌鲁木齐830011 [2]新疆喀什地区第一人民医院肿瘤科,新疆喀什844000

出　　处：《新疆医科大学学报》2018年第8期987-990,共4页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金(2016D01C376)

摘　　要：目的探索人非小细胞肺癌血管内皮细胞(HNSCLC-MECs)的培养方法,观察细胞生长状态并鉴定。方法收集外科非小细胞肺癌手术组织标本,研磨至匀浆经钢筛网过滤收集沉淀,短时少量胰酶消化后组织块贴服培养。根据细胞差速粘附的原理对细胞进行纯化和传代。结果原代培养60h后可见少量细胞贴壁生长,7d后贴壁细胞明显增多,传代后10d单层细胞呈典型"铺路石"样改变。应用激光共聚焦显微镜可见FactorⅧ及CD34抗体表达于细胞膜上,表达阳性率>95%。结论应用筛网法、贴壁法和少量胰酶法可以获得大量生长良好的HNSCLC-MECs,经鉴定符合血管内皮细胞特性。Objective To explore a cultivation method of microvascular endothelial cells in human Nonsmall lung cancer.Methods Collected the Non-small lung cancer tissue from surgical operation,ground the tissue to homogenate and filtered by microscreen to collect precipitation.Human Non-small lung cancer-microvascular endothelial cells（HNSCLC-MECs）were obtained by a small amount of short-term trypsin method,followed by purified utilizing differential adhesion method when passage culture.ResultsPrimary cultured endothelial cells completely adhered to the wall at 60 hand increased after 7 d later,after 10 dpassage,the HNSCLC-MECs displayed typical cobblestone-like morphology.Laser scanning confocal microscopy showed that cultured cells were positive for FVIII and CD34-associated antigens with 95% positive rate.Conclusion These results suggest that abundant highly purified HNSCLC-MECs can be obtained through the filter method,adherent method and a small amount of short-term trypsin method and proved to be in line with the characteristics of endothelial cells.

关 键 词：人非小细胞肺癌血管内皮细胞 原代培养 细胞鉴定 

分 类 号：R730.231[医药卫生—肿瘤]