与成纤维细胞共培养的肺泡II型上皮细胞的生物学特性

Biological Characteristics of Type II Alveolar Epithelial Cells Co-cultured with Lung Fibroblasts

建立胎鼠肺泡II型上皮细胞(AECII)与肺成纤维细胞(LF)共培养模型,观察与LF共培养下AECII的生物学特性。倒置相差显微镜观察AECII形态和基本生长情况;RT-PCR和流式细胞术分别检测肺泡表面活性蛋白-C(SP-C)、水通道蛋白5(AQP5)mRNA及蛋白质表达;流式细胞术检测细胞周期及Ki67表达。结果显示,与LF共培养时,AECII能较好地保留其细胞形态,SP-CmRNA及其蛋白质表达明显增加,而AQP5mRNA及其蛋白质表达则明显减少;LF促进AECII增殖,使G2/M、S期细胞及表达Ki67+细胞的比率明显增多。结果提示,AECII与LF共培养时,能更好地保留其细胞形态、分化及增殖特性。

To observe the biological characteristics of type II alveolar epithelial cells （AECⅡ） co-cultured with lung fibroblasts （LF） in vitro, a co-culture model of AECⅡ and LF from fetal rat lung was set up. The morphological change and growth of AECⅡ were observed under inverted phase contrast microscope. The SP-C and AQP5 expressions in mRNA and protein levels were observed by RT-PCR and flow cytometry. Flow cytometry was also used to detect the cell cycle and Ki67 expression of AECⅡ. When co-cultured with LF, AECII could keep its morphology well, and the SP-C expression in mRNA and protein levels increased markedly, while the AQP5 expression decreased. LF facilitated the proliferation of AECII by increased the percentage of cells in S and G2/M phases and the expression rate of Ki67. The results showed that AECⅡ can keep its morphology, differentiated and proliferation characteristics when co-cultured with LF.