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Isolation of a putative progenitor subpopulation of alveolar epithelial type 2 cells.

机译:肺泡上皮2型细胞的推定祖细胞亚群的分离。

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摘要

Alveolar epithelial type 2 cells (AEC2) isolated from hyperoxia-treated animals exhibit increases in both proliferation and DNA damage in response to culture. AEC2 express the zonula adherens proteins E-cadherin, -, - and -catenin, desmoglein, and pp120, as demonstrated by Western blotting. Immunohistochemical analysis of cultured AEC2 showed expression of E-cadherin on cytoplasmic membranes varying from strongly to weakly staining. When cultured AEC2 placed in suspension were labeled with fluorescent-tagged antibodies to E-cadherin, cells could be sorted into at least two subpopulations, either dim or brightly staining for this marker. With the use of antibody to E-cadherin bound to magnetic beads, cells were physically separated into E-cadherin-positive and -negative subpopulations, which were then analyzed for differences in proliferation and DNA damage. The E-cadherin-positive subpopulation contained the majority of damaged cells, was quiescent, and expressed low levels of telomerase activity, whereas the E-cadherin-negative subpopulation was undamaged, proliferative, and expressed high levels of telomerase activity.
机译:从高氧血症治疗的动物中分离出的肺泡上皮2型细胞(AEC2)在培养过程中表现出增殖和DNA损伤增加。如Western blotting所示,AEC2表达小带粘附蛋白E-cadherin,-,-和-catenin,desmoglein和pp120。培养的AEC2的免疫组织化学分析显示,E-钙粘着蛋白在细胞膜上的表达从强到弱染色不等。当将悬浮的培养的AEC2用抗E-cadherin的荧光标记抗体标记时,可以将细胞分为至少两个亚群,该标记为暗染或亮染。使用结合到磁珠的E-钙粘蛋白抗体,将细胞物理分离为E-钙粘蛋白阳性和阴性亚群,然后分析其增殖和DNA损伤的差异。 E-cadherin阳性亚群包含大部分受损细胞,处于静止状态,并表达低水平的端粒酶活性,而E-cadherin阴性亚群则未受损,增殖并表达高水平的端粒酶活性。

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