关键词: 根尖牙乳头干细胞, 成神经分化, WDR63, 类神经球, 神经分化指标

Abstract: Objective：To investigate the effect of dynein 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: Using lentiviral 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. 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. 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 and protein expression (P＜0.001), significant increase in neuronal ball diameter (P＜0.01), and obvious increased in Nestin and βIII-tubulin-positive neural ball-like cells with significantly enhanced fluorescence intensity (P＜0.01). βⅢ-tubulin, NeuroD, NCAM, and Nestin gene expression also increased (P＜0.05). Conversely, knockdown of WDR63 led to decreased WDR63 gene and protein expression (P＜0.01), 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.

Key words: stem cells from the apical papilla, differentiation into neurons, WDR63, neurospheres, neural differentiation markers