WDR63 promotes the neural differentiation potential of human apical papilla stem cells
Objective:To investigate the effect of WDR63 on the differentiation ability of human stem cells from the apical papilla(SCAPs)and the expression of neural differentiation-related markers in vitro.Methods:qRT-PCR was used to detect the expression changes of WDR63 gene in untransfected lentivirus SCAPs at 3,6 and 9 days after induction of neurogenic differentiation.Using lenti-viral transfection,stable over-expression and knockdown of WDR63 in SCAPs were obtained.The neural differentiation of the cells was induced through a neurosphere formation experiment,and the diameter of the neurospheres was quantitatively analyzed.The expression of Nestin and βⅢ-tubulin in SCAPs was detected by immunofluorescence staining and quantitatively analyzed.qRT-PCR was used to detect the expression of neural differentiation-related indicators such as βⅢ-tubulin,NeuroD,NCAM,and Nestin genes.Results:The expression level of WDR63 in untransfected lentivirus SCAPs gradually increased at 3,6,and 9 days after neuronal differentiation(P＜0.05).Overexpression of WDR63 led to the increase of WDR63 gene(P＜0.001)and protein expression,significant increase in neuronal ball diameter(P＜0.01),and obvious increased in Nestin and βⅢ-tubulin-positive neural ball-like cells with significantly en-hanced fluorescence intensity(P＜0.01).βⅢ-tubulin,NeuroD,NCAM,and Nestin gene expression also increased(P＜0.05).Con-versely,knockdown of WDR63 led to decrease WDR63 gene(P＜0.01)and protein expression,significant reduction in neuronal ball diameter(P＜0.05),and significant decrease in Nestin and βⅢ-tubulin-positive neural ball-like cells(P＜0.05)with decreased βⅢ-tubulin,NeuroD,NCAM,and Nestin gene expression(P＜0.05).Conclusions:WDR63 promotes the neural differentiation potential of SCAPs.
stem cells from the apical papilladifferentiation into neuronsWDR63neurospheresneural differentiation markers
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