Neural Crest Stem Cells from Dental Tissues: A New Hope for Dental and Neural Regeneration
Dental EMSCs express neural differentiation and pluripotency markers and can acquire a prominent neural-like morphology in vitro. Dental EMSCs isolated from dental pulp (DPSCs) form clonogenic adherent colonies (a), which present Nestin+ immunoreactivity (b) and from which equally Nestin+ migrating cells spread to eventually bring the culture plate to full confluency (c). Dental EMSCs also express pluripotency markers such as Oct-4, in the absence of any genetic or pharmacological manipulation (d). The cellular morphology and proliferation rates of dental EMSCs vary depending on the presence of FBS in the culture medium. DPSCs proliferate slowly in the absence of serum. Cells cultured without serum are equally Nestin+ but display very variable morphologies, including the appearance of cells with striking neuron-like shape, that show very thin and long cytoplasmic processes, resembling dendrites and axons (f, g). When DPSC are expanded for 3 weeks with 10% FBS, following another 3 weeks of serum deprivation, a sheet of nerve-like tissue is formed (h). Times after seeding: (a) 1 week; (e) 3 weeks; (h) 6 weeks (3 + 3); (b–d; f–g) double merged images of Nestin (green) and Oct-4 (red) immunolabeled cells, with DAPI (blue) introduced as a nuclear counterstain. Scale bars: 50 m.