肺泡Ⅱ型上皮细胞的分离培养和磷脂酶A_2的损伤作用  

Isolation and Culture of Alveolar Type Ⅱ Epithelial Cells and Injurious Effects of Phospholipase A_2 on the Cultured Cells

作　　者：丁自强[1] 吴中立[1] 

机构地区：[1]第二军医大学训练部病理生理学教研室,200433

出　　处：《第二军医大学学报》1992年第1期17-21,共5页Academic Journal of Second Military Medical University

摘　　要：本文报道了豚鼠肺泡Ⅱ型上皮细胞的分离和原代培养方法,并在体外研究了磷脂酶A_2(PLA_2)对培养细胞的损伤作用。将胰酶消化液注入支气管肺泡腔,肺组织消化后剪碎、振摇、过滤,可得到172.3×10~6细胞/豚鼠,Ⅱ型细胞占33.6％。细胞经短期贴壁培养选择,再连续培养48h,可得到7.9×10~6细胞/豚鼠,Ⅱ型细胞占66.1％,活力96.2％。贴壁细胞经EDTA处理,Ⅱ型细胞纯度提高到76.2％(63.9％～92％)。培养细胞经相差显微镜、光镜和电镜观察证实为Ⅱ型细胞,在培养液中加PLA_2 300U/ml作用4h,细胞脱落率明显增高,乳酸脱氢酶释放增加,说明PLA_2对细胞有损伤和促使脱落的作用,可能在肺损伤发生中有重要意义。A method for isolation and primary culture of guinea pig alveolar type Ⅱ epithelial cells is reported. The injurious effects of phospholipase A2 (PLA2) on the cultured cells were studied. Trypsinizing solution was instilled into the bronchoalveolar space. The lungs were digested, sectioned, vortexed and filtered. The yield of original cell mixture was 172.3×106 cells/guinea pig, of which 33.6% were type Ⅱcells. At 48 h of culture following short term selective attachment 7.9×106 cells were obtained, of which 66.1% were type Ⅱ ceils. The purity could further be raised to 76.2% (63.9% -92%) if pretreated with ethyienediarnine tetraacetic acid (EDTA) before trypsinization to remove attached cells. The characteristics of cultured cells were observed under phase contrast microscopy, light microscopy and electron microscopy. After exposure of the cells to 300U/ml PLA2 for 4h at 37℃, detachment and lactic dehydrogenase (LDH) release of the cultured cells increased significantly. It is suggested that PLA, may damage and promote detachment of cultured type Ⅱ cells and it may play an important part in the development of some diseases.

关 键 词：肺泡 细胞培养 细胞分离 磷脂酶A 

分 类 号：R563.02[医药卫生—呼吸系统]