Daucosterol promotes the proliferation of neural stem cells

Abstract

Neural stem cells (NSCs) are self-regenerating cells, but their regenerative capacity is limited. The present study was conducted to investigate the effect of daucosterol (a sterolin) on the promotion of NSC proliferation and determine the corresponding molecular mechanism. Results of cell counting kit-8 (CCK-8) assay showed that daucosterol significantly increased the quantity of viable cells and the effectiveness of daucosterol was similar to that of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). Flow cytometry detection of CFSE-labeled (CFSE, carboxyfluorescein diacetate succinimidyl ester) NSCs showed that Div Index (or the average number of cell divisions) and % Divided (or the percentage of cells that divided at least once) of the cells were increased, indicating that daucosterol increased the percentage of NSCs re-entering the cell cycle. mRNA microarray analysis showed that 333 genes that are mostly involved in the mitotic cell cycle were up-regulated. By contrast, 627 genes that are mostly involved in differentiation were down-regulated. In particular, insulin-like growth factor I (IGF1) was considered as an important regulatory gene that functionally promoted NSC proliferation, and the increased expression of IGF1 protein was validated by ELISA. In addition, the phosphorylation of AKT was increased, indicating that the proliferation-enhancing activity of daucosterol may be involved in IGF1-AKT pathway. Our study provided information about daucosterol as an efficient and inexpensive growth factor alternative that could be used in clinical medicine and research applications.

Keywords: (8)%D or % Divided; 4’ 6-diamidino-2-phenylindole; BCA; Bicinchoninic Acid; CCK-8; CFSE; DAPI; DI or Div Index; Daucosterol; EGF; FITC; GO; Gene Ontology; IGF1; NSCs; Neural stem cells; Proliferation; bFGF; basic fibroblast growth factor; carboxyfluorescein diacetate succinimidyl ester; cell counting kit-8; epidermal growth factor; fluorescein isothiocyanate; fold change; insulin-like growth factor I; the average number of divisions; the percentage of the cells of the original sample which divided at least once; the ratio of normalized intensities between the two conditions.