Improved primary culture and primary identification of human pituitary adenoma cells

Abstract

Objective To explore an improving primary culture method of human pituitary adenoma cells. Methods Thirty-six pituitary adenoma specimens, collected from excision and conformed by pathology in our hospital from November 2014 to June 2015, were used. Based on the former experience from papers and ourselves, our primary culture methods of several different kinds of human pituitary adenomas were improved. Purification and passage culture of cells were performed in the following experiments. CCK-8 assay was used to detect the multiplication capacity of pituitary adenoma cells; immunofluorescent staining and Western blotting were used to observe the cytokeratin and vimentin protein expressions. Results By using the improve culture methods, tumor cells proliferated in suspension with good status and had the tendency of aggregation in culture medium. The proliferation test showed that the tumor cells enjoyed proliferation ability in vitro; tumor cells could pass over five generations. The growth curve of pituitary adenoma cells showed S pattern. Immunofluorescent staining and Western blotting indicated negative cytokeratin and vimentin protein expressions, showing that the cells were from epithelium. Conclusion Each type of pituitary adenoma cells has characteristics of suspended growth and tendency of gathering by using this improved method. Key words: Pituitary adenoma; Cell culture; Cell line