人气道平滑肌原代细胞不同提取方法对比  

Comparison of primary cells extraction methods of human airway smooth muscle

作　　者：黄兰芳 黄青华 李梦泽 侯长春 闫萍[1] Huang Lanfang;Huang Qinghua;Li Mengze(The First Affiliated Hospital of Guangxi Medical University,Nanning 530021)

机构地区：[1]广西医科大学第一附属医院老年呼吸内科,广西南宁530021 [2]广西医科大学第一附属医院呼吸与危重症学科,广西南宁530021 [3]广西医科大学第二附属医院

出　　处：《中国现代医药杂志》2020年第4期1-3,共3页Modern Medicine Journal of China

基　　金：国家自然科学基金资助项目(编号:81600020)。

摘　　要：目的对比人气道平滑肌原代细胞不同提取的方法,探讨各个方法的利弊,为后续细胞实验提供基础。方法收集我院胸外科2019年3~10月行肺叶切除术肺癌患者的正常肺叶、段支气管标本15例,采用酶解离法和组织块贴壁法提取气道平滑肌细胞,倒置相差显微镜观察其生长情况,用免疫荧光法检验平滑肌中特异性α-actin的表达。结果使用组织块贴壁法和酶解离法分别进行细胞原代培养,15例均成功,成功率100%,污染率为0。组织块贴壁法平均5~7天可见细胞贴壁生长,15~20天出现融合;酶解离法平均7~10天可见细胞增多融合。传代后细胞呈“峰-谷”生长。免疫荧光法鉴定气道平滑肌细胞纯度达到95%。结论两种方法都能提取人气道平滑肌原代细胞并进行后续培养,组织块贴壁法对细胞活力影响小,且经济方便,值得推广使用。Objective To compare the primary cells extraction methods of human airway smooth muscle,discuss the advantages and disadvantages of each method,and to provide a basis for subsequent cell experiments.Methods From Mar to Oct in 2019,15 normal pulmonary lobe and segment bronchus specimens of patients undergoing lobectomy for lung cancer in the department of thoracic surgery of our hospital were extracted by enzymatic digestion and tissue block attachment method,and the growth of airway smooth muscle cells was observed by inverted phase contrast microscope.Results 15 cases of primary cell culture using tissue block attachment method and enzymatic method were successful,with a success rate of 100%,the pollution rate was 0.On average,cell adherent growth was observed in 5~7 days by tissue block attachment method,and fusion occurred in 15~20 days.Increased cell fusion was observed on average 7~10 days by enzymatic method.After transplanting,the cells showed a“peak-valley”growth.The purity of airway smooth muscle cells was 95%.Conclusion Both methods can extract primary cells of human airway smooth muscle and conduct subsequent culture.Tissue block attachment method has little effect on cell viability and is economical and convenient.

关 键 词：人气道平滑肌细胞 组织块贴壁法 酶解离法 

分 类 号：R562.25[医药卫生—呼吸系统]