Objective To explore the application of tissue-explant technique in culturing rat submandibular gland cell(RSGC)and the characteristics of RSGC.Methods RSGCs were cultured using tissue-explant technique.The cells were purified by enzymatic digestion and differential adhesion.The cell phenotype was identified by cytokeratin-8 immunohistochemical staining.Cellular morphology was observed and photographed under inverted microscope.The cell viability and growth were determined by a double-staining procedure using FDA-PI and MTT assay,respectively.Results Cytokeratin-8 was positive stained in the immunohistochemical staining.The cell viability was more than 95%.The cell growth curve showed that RSGCs were in logarithmic phase since day 5.Conclusion Tissue-explant technique is an easy way to purify plentiful RSGC with normal functions,and it can be used in further research of tissue engineering of submandibular gland.
Result Analysis
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