Experimental Study On Epithelial Differentiation Of ADSCs Induced By Saliva

Objective: To investigate the differentiation of rabbit adipose stem cells(ADSCs)induced by Saliva in vitro and observe whether it could promote their epithelial differentiation.Methods: Fat harvested from 2-3 months New Zealand White Rabbit was isolated,cultured,generated ADSCs.The experiment was divided into three groups: ADSCs group,ADSCs+EGF group and ADSCs+Saliva group.ADSCs was induced by EGF(10 ng/ml)and saliva(15% concentration)in the induced group,while ADSCs group(non-induced group)served as negative control group.Observed the cell morphology changes of ADSCs induced by saliva,and detected cell proliferation status by CCK8 method.The expression of CK19 was detected by immunofluorescence,RT-PCR and WB respectively after 7 days and 14 days of induction.Results:Under the microscope,it was shown that the cell morphology of ADSCs with saliva was gradually changed from long spindle shape to short spindle shape or even irregular oblate shape.In the 7 and 14 days EGF and saliva induction groups,the cell epidermal marker CK19 was positively expressed in immunofluorescence detection.Although the expression in saliva induction group was not as strong as that in EGF induction group,it was also higher than that in negative control group,and the expression level of CK19 in the 14-day induction group was significantly higher than that in the 7-day induction group.The fluorescence quantitative PCR showed that the expression of CK19 mRNA in EGF and saliva induced group was higher than that in negative control group(P<0.01).And the expression level at 14 days was significantly higher than that at 7 days(P<0.01).For the detection of protein level in each group,the results were consistent with the expression level of mRNA.Conclusion: Saliva could promote the epithelial differentiation of ADSCs to some extent and induce its epithelial transformation.