大鼠肺泡II型上皮细胞系RAE-1的建立及其生物学特性

建立肺泡II型细胞系为研究细胞恶性转化提供实验模型。采用Nycodenz密度梯度离心法分离了大鼠肺泡II型上皮细胞 ,原代培养大鼠肺泡II型上皮细胞并将SV4 0病毒的早期区大T基因转导入上皮细胞 ,建立了可在体外长期稳定传代的细胞系RAE - 1。对此细胞系的生物学特性分析表明 :细胞的基因组中整合了SV4 0T基因 ;染色体分析以非整倍体为主 ,众数染色体为 4 4条 ;细胞不能在软琼脂上形成克隆 ;除了转化特性以外 ,RAE - 1细胞仍保留了正常的上皮组织角蛋白Ck8、Ck18的表达

Establishment of rat alveolar type II epithelial cell line RAE-1 and its biological characteristics

The aim is to establish rat alveolar type II epithelial cell line and to provide cell models for epithelial cell cancer research. Highly pure Lewis rat alveolar epithelial cells were successfully isolated by Nycodenz buoyant density gradient centrifugation in a combination with IgG panning method. Semian virus 40 early region large T gene was virally transferred into primary cultures. After neo gene selection, a cell line RAE-1 was established. The characteristics of this cell line were analyzed. Homologous integration of SV40T gene into RAE-1 cell genome was positively detected by PCR, modal chromosome number of RAE-1 cells was 44, and expressions of cytokeratins 8 and 18 at mRNA level were both positively detected by RT-PCR. The establishment of RAE-1 cell line was very important because it could serve as model cells to study epithelial cell neoplastic transformation mechanisms in hazards-caused alveolar epithelial cancer.