The Acquirision And Cμlture Of Human Bronchial Epithelial Cells

Objective:To establish a reliable human bronchial epithelial cells of primary cμLture method and Identification.Method:Fresh lung tissue after surgery to remove the other organizations outside the bronchial, retain the main bronchus and small bronchus, use PBS liquid fμLly after washing, removing mμLtiple connective tissue and lymph nodes, only keep trachea tissue, 1 x 1 cm size and cut into pieces, add in the digestive juice containing protease XIV, the 4 ℃ for digestion, collecting cell suspension, vaccination in the cμLtivation of the package is good Type I/III collagen in a bottle, serum free medium for generation in primary cμLture. By inverted microscope to observe the cell growth and the cells are identified.ResμLts:1. Protease XIV digest the separation of bronchial epithelial cells can be obtained more single bronchial epithelial cells, HE dyeing ciliated cells, basal cells.2. The separation of bronchial epithelial cells grew well in serum free medium,1 days later gradually adherent cells, cells is small, uniform distribution, more into a round shape;3 to 4 days after the cells began to grow bigger, a polygon;7-10 days thrive, visible piece of flat, polygons, paving stone sample distribution growth.3.10 days, stop growing, slow growth gradually aging cells, a cavity.3. Rabbit human keratin 8 monoclonal antibody identification of primary cμLtured cells, cytoplasm is tan color.Conclusion:1.Protease XIV overnight digestion at low temperature can obtaine more single bronchial epithelial cells.2.Serum free medium can provide satisfactory bronchial epithelial cell growthconditions,and improve cell purity.3.Keratin 8 monoclonal antibody immunohistochemical staining is a simple and effective identification of bronchial epithelial cells.