摘要

实验于2007-10/12在北京军区总医院附属八一儿童医院实验室完成。取孕19 d SD大鼠胎鼠肺组织进行原代培养，通过IgG包被瓶培养进行筛选纯化。分别于培养24，48，72 h 后检测细胞的活力；利用共聚焦显微镜及电镜对肺泡Ⅱ型上皮细胞进行鉴定。各时间点Ⅱ型细胞活力均在90%以上，细胞纯度在91%以上，电镜下细胞胞浆内含有丰富的板层小体，细胞表面有微绒毛；共聚焦显微镜下细胞浆中有呈绿色荧光的SP-C阳性表达。胰蛋白酶加胶原酶消化，IgG包被瓶进行筛选所得肺泡Ⅱ型细胞活力及纯度均较高，可以作为肺泡Ⅱ型细胞的原代培养的常规方法；电镜和免疫荧光均可达到鉴定目的。
关键词：肺泡Ⅱ型细胞；原代培养；鉴定；组织构建

刘秀香，杨志军，陈洪清，封志纯.胎鼠肺泡Ⅱ型细胞的原代培养及鉴定[J].中国组织工程研究与临床康复，2008，12(18):3497-3499 [www.zglckf.com/zglckf/ejournal/upfiles/08-18/18k-3497(ps).pdf]

Primary culture and identification of type Ⅱ alveolar epithelial cells from fetal rats

Abstract

The experiment was performed in Bayi Children's Hospital, General Hospital of Beijing Military Area Command of Chinese PLA from October to December 2007. Lung tissues of fetal rats of SD gestation rats for 19 days were cultured primarily and purified by culture flasks coated with rat IgG. Cell viability was detected at 24, 48 and 72 hours of culture. Type Ⅱ alveolar epithelial cells (AEC Ⅱ) were identified of confocal microscopy and electron microscope. On every time point, the viability of AEC Ⅱ was above 90%, and the purity was above 91%. The cells had plenty of lamellar bodies in cytoplasm and many microvilli on cell surface; there was green fluorescence expression of SP-C in cytoplasm. The viability and purity of AEC Ⅱobtained by digestion of trypsin and collagenase and by screening with culture flasks coated with rat IgG is higher. Therefore, this method can serve as a routine method of primary culture of AEC Ⅱ. Both electron microscope and immunofluorescence can identify the cells.

Liu XX, Yang ZJ, Chen HQ, Feng ZC.Primary culture and identification of type Ⅱ alveolar epithelial cells from fetal rats.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(18):3497-3499
[www.zglckf.com/zglckf/ejournal/upfiles/08-18/18k-3497(ps).pdf]