鸡胚原代肺泡 II 型上皮细胞的分离培养与鉴定.

[Objectives] The aim of the study was to establish a stable technique for isolation, purification and culture of chicken type II alveolar epithelial cells, and to provide an in vitro research model for investigation of the effects of air pollutants and other environmental stress factors on chicken alveolar function. [Methods] Digestion of lung tissue from 16-day-old chicken embryos was conducted with type I collagenase. After differential centrifugation, and differential attachment, chicken immunoglobulin G (lgG) immunoadsorption purification of cells was performed, and immunofluorescence detection and alkaline phosphatase were used for cell identification. [Results] The cells were adhered for 18-24 h, and they were polygonal in shape and grew like islands. After 24-72 h, the cells had proliferated and metabolized vigorously, the nucleus was obvious, the cytoplasm was rich, and the cells gradually connected to form a monolayer. After 72 h, the intracellular particulate matter decreased and the cell morphology changed. lmmuno-fluorescence detection showed the expression of epithelial cell marker cytokeratin-19(CK19). Alkaline phosphatase staining showed diffuse red or red-brown particles in the cytoplasm. [Conclusions] This method is an effective technique for the isolation, purification and culture of chicken alveolar epithelial cells with high cell yield and purity, which can satisfy general research in vitro. [ABSTRACT FROM AUTHOR]