In Vitro Study On Differentiation Of Canine Bone Marrow Mesenchymal Stem Cells Into Epithelium-like Cells

Objective To investigate the feasibility and basic conditions of canine bone marrow mesenchymal stem cells(BMSCs)differentiated into epithelium-like cells in vitro. Methods BMSCs were isolated and purified by density gradient centrifugation and anchoring culture. After proliferation BMSCs were treated in induced mediums and direct-contact cultivated with canine esophageal epithelial cells(EECs).The morphology of BMSCs was examined everyday by inverted phase contrast microscope.Cultured cells were identified by immunofluorescence assay, immunocytochemistry staining and flow cytometry at different time to detect the positive staining of pan-cytokeratin(CK) and epithelial membrane antigen(EMA). Results The highly purified BMSCs possessed the capabilities of rapid proliferation and pluripotential differentiation. The CK or EMA positive cells could be observed after Serum-free Keratinocyte Medium(K-SFM) consisting of 20ng/ml epidermal growth factor(EGF) treatment. The number of positive cells was correlated to the density of EGF and induced time.BMSCs could be induced into epithelium-like cells after coculturing with EECs. Conclusion Our experiment indicates canine BMSCs can differentiate into epithelium-like cells in vitro. As a result, BMSCs will serve as seed cells for tissue engineered esophagus in the future.