Small molecules enhance neurogenic differentiation of dental-derived adult stem cells.
OBJECTIVE
Dental-derived stem cells originate from the embryonic neural crest, and exhibit high neurogenic potential. This study aimed to investigate whether a cocktail of eight small molecules (Valproic acid, CHIR99021, Repsox, Forskolin, SP600125, GO6983, Y-27632 and Dorsomorphin) can enhance the in vitro neurogenic differentiation of dental pulp stem cells (DPSCs), stem cells from apical papilla (SCAPs) and gingival mesenchymal stem cells (GMSCs), as a preliminary step towards clinical applications.


MATERIALS AND METHODS
Neural induction was carried out with a small molecule cocktail based two-step culture protocol, over a total duration of 14 days. At the 8 and 14 day timepoints, the cells were analyzed for expression of neural markers with immunocytochemistry, qRT-PCR and Western Blot. The Fluo 4-AM calcium flux assay was also performed after a further 14 days of neural maturation.


RESULTS
More pronounced morphological changes characteristic of the neural lineage (i.e. neuritogenesis) were observed in all three cell types treated with small molecules, as compared to the untreated controls. This was corroborated by the immunocytochemistry, qRT-PCR and western blot data, which showed upregulated expression of several early and mature neural markers in all three cell types treated with small molecules, versus the corresponding untreated controls. Finally, the Fluo-4 AM calcium flux assay showed consistently higher calcium transient (F/Fo) peaks for the small molecule-treated versus untreated control groups.


CONCLUSIONS
Small molecules can enhance the neurogenic differentiation of DPSCs, SCAPs and GMSCs, which offer much potential for therapeutic applications.
['Heng|Boon Chin|BC|', 'Jiang|Shan|S|', 'Yi|Baicheng|B|', 'Gong|Ting|T|', 'Lim|Lee Wei|LW|', 'Zhang|Chengfei|C|']
D000328:Adult D053687:Adult Stem Cells D002454:Cell Differentiation D002478:Cells, Cultured D003782:Dental Pulp D006801:Humans D055495:Neurogenesis D013234:Stem Cells
Differentiation Stem cells DPSCs SCAPs GMSCs Neurogenesis