Experiment of oral mucosa epithelial cells cultured on small intestinal submucosa in vitro

Objective To explore an effective method to culture oral mucosa epithelial cells(OMECs) of canine in vitro,and to observe the biological characteristics of OMECs growing on small intestinal submucosa(SIS) in order to provide the experimental basis for epithelium tissue engineering.Methods The primary OMECs were cultivated with DKSFM(defined keratinocyte serum free medium) containing 6% fetal bovine serum(FBS).The morphological characteristics and the growth curve of OMECs were observed.The expressions of OMECs marker(CK19) were examined by immunocytochemistry.The 2nd passage of OMECs were seeded on SIS,OMECs co-cultured with SIS were observed by hematoxylin-eosin staining,immunohistochemical staining,and scanning electron microscope(SEM).Results OMECs were grown well in DKSFM.Immunohistochemical staining of the 2nd passage cultured canine OMECs with broadly reacting anti-cytokeratin anyibodies(CK19) was positive.OMECs formed a single layer on the surface of SIS,and eight days later the cells were polygong and arranged like slabstone.Conclusion Culture of canine OMECs in DKSFM containing 6% FBS is a simple and feasible method.SIS has good biocompatibility,it is a kind of good bioscafold in the tissue-engineered epithelium.