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Braz. j. med. biol. res ; 51(2): e6950, 2018. tab, graf
Article Dans Anglais | LILACS | ID: biblio-889028

Résumé

Alveolar epithelia play an essential role in maintaining the integrity and homeostasis of lungs, in which alveolar epithelial type II cells (AECII) are a cell type with stem cell potential for epithelial injury repair and regeneration. However, mechanisms behind the physiological and pathological roles of alveolar epithelia in human lungs remain largely unknown, partially owing to the difficulty of isolation and culture of primary human AECII cells. In the present study, we aimed to characterize alveolar epithelia generated from A549 lung adenocarcinoma cells that were cultured in an air-liquid interface (ALI) state. Morphological analysis demonstrated that A549 cells could reconstitute epithelial layers in ALI cultures as evaluated by histochemistry staining and electronic microscopy. Immunofluorescent staining further revealed an expression of alveolar epithelial type I cell (AECI) markers aquaporin-5 protein (AQP-5), and AECII cell marker surfactant protein C (SPC) in subpopulations of ALI cultured cells. Importantly, molecular analysis further revealed the expression of AQP-5, SPC, thyroid transcription factor-1, zonula occludens-1 and Mucin 5B in A549 ALI cultures as determined by both immunoblotting and quantitative RT-PCR assay. These results suggest that the ALI culture of A549 cells can partially mimic the property of alveolar epithelia, which may be a feasible and alternative model for investigating roles and mechanisms of alveolar epithelia in vitro.


Sujets)
Humains , Milieux de culture conditionnés , Techniques de culture cellulaire/méthodes , Pneumocytes/physiologie , Cellules A549/physiologie , Valeurs de référence , Facteurs temps , Microscopie électronique à balayage , Immunotransfert , Numération cellulaire , Reproductibilité des résultats , Analyse de variance , Protéine C associée au surfactant pulmonaire/analyse , Aquaporine-5/analyse , Mucine 5B/analyse , Réaction de polymérisation en chaine en temps réel , Protéine-1 de la zonula occludens/analyse , Facteur-1 de transcription de la thyroïde/analyse
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