小鼠胎鼠肺泡Ⅱ型上皮细胞的分离、原代培养及鉴定  

Isolation,Primary Culture and Identification of Type Ⅱ Alveolar Epithelial Cells from Fetal Mouse

作　　者：陈坤[1] 邓世雄[1] 刘芳君[2] 李虹[3] 蒋朴[1] 

机构地区：[1]重庆医科大学基础医学院法医学教研室,重庆400016 [2]重庆医科大学附属儿童医院,儿童发育疾病研究省部共建教育部重点实验室,重庆400014 [3]重庆市公安局江北分局,重庆400020

出　　处：《西南大学学报（自然科学版）》2011年第12期62-65,共4页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(30772459)

摘　　要：目的:建立系统可靠的小鼠胎鼠肺泡Ⅱ型上皮细胞的分离、培养、纯化及鉴定技术,为进一步研究肺纤维化及肺肿瘤等疾病的发病机制奠定基础.方法:应用低质量分数胰酶联合胶原酶的方法消化小鼠胎鼠肺组织块,经差速离心和差速贴壁的方法纯化肺泡Ⅱ型上皮,进行原代培养.用改良巴氏染色法和透射电镜观察对所分离的细胞进行鉴定.结果:每只胎鼠可获得(5±2)×106的细胞产量,细胞活力为95%±2%,纯度达到92%±2%,改良巴氏法镜下观察可见胞质内含较多深色颗粒,透射电镜观察到特征性的嗜锇小体即板层小体和细胞膜上有明显的微绒毛.结论:该方法能成功分离出小鼠胎鼠肺泡Ⅱ型上皮细胞,且产量大、纯度高,能满足体外研究的需要.Objective： To develop a reliable method for the isolation,purification,primary culture and identification of AEC Ⅱ（type Ⅱ alveolar epithelial cells） from the fetal mouse lung and lay a foundation for the further study of pulmonary fibrosis and lung cancer pathogenesis.Methods： Lung tissues of fetal mice were digested with low-density trypsin and collagenase,and then AECⅡwere purified with different centrifugal force and repeated attachment for primary culture.The isolated cells were identified by transmission electron microscopy（TEM） and modified Papanicolaou dying.Results： An average of（5±2）×106 cells were obtained from each fetal mouse,their vitality was 95% ±2% and their purity was 92%±2%.Many granules were observed within the cytoplasm by modified Papanieolaou dying,and [osmiophilic] lamellar bodies and microvilli were found by electron microscopy.Conclusion： This method can successfully produce a large number of highly purified ACEⅡ from the fetal mouse lung,which serves the needs in the study in vitro.

关 键 词：胎鼠 肺泡Ⅱ型上皮细胞 原代培养 鉴定 

分 类 号：Q813.1[生物学—生物工程]