Abstract
Dental pulp stem cells (DPSCs) have the capacity to give rise to cells with neuronal-like phenotypes, suggesting their use in brain cell therapies. In the present work, we wanted to address the phenotypic fate of adult genetically unmodified human DPSCs cultured in NeurocultTM (Stem Cell Technologies), a cell culture medium without serum which can be alternatively supplemented for the expansion and/or differentiation of adult neural stem cells (NSCs). Our results show that non-genetically modified human adult DPSCs cultured with Neurocult NS-A proliferation supplement generated neurosphere-like dentospheres expressing the NSC markers Nestin and glial fibrillary acidic protein (GFAP), but also the vascular endothelial cell marker CD31. Remarkably, 1 month after intracranial graft into athymic nude mice, human CD31+/CD146+ and Nestin+ DPSC-derived cells were found tightly associated with both the endothelial and pericyte layers of brain vasculature, forming full blood vessels of human origin which showed an increased laminin staining. These results are the first demonstration that DPSC-derived cells contributed to the generation of neovasculature within brain tissue, and that Neurocult and other related serum-free cell culture media may constitute a fast and efficient way to obtain endothelial cells from human DPSCs.
Highlights
The dental pulp is home to an active population of neural crest stem cells, which are responsible of renewing adult differentiated cells, such as odontoblastic cells and Schwann cells, within the pulp tissue (Kaukua et al, 2014; Aurrekoetxea et al, 2015)
Previous research had shown that the dental pulp tissue of adult teeth contained neural-crest derived dental pulp stem cells (DPSCs), which expressed neural stem cells (NSCs) markers such as Nestin and glial fibrillary acidic protein (GFAP) (Gronthos et al, 2000; Chang et al, 2014)
This is the first time in which DPSCs, disaggregated from floating dentospheres grown in a serumfree medium and grafted into the brain of immunosuppressed mice, give rise to CD31-positive endothelial and CD146-positive pericyte cells that can integrate into the host brain vasculature, promoting de novo generation of new blood vessels containing both endothelial and mural layers, within brain tissue
Summary
The dental pulp is home to an active population of neural crest stem cells, which are responsible of renewing adult differentiated cells, such as odontoblastic cells and Schwann cells, within the pulp tissue (Kaukua et al, 2014; Aurrekoetxea et al, 2015). These cells are referred to as dental pulp stem cells (DPSCs) when extracted from permanent teeth, and as stem cells from human exfoliated deciduous teeth (SHEDs) when extracted from first generation teeth during childhood (Gronthos et al, 2000, 2002; Miura et al, 2003). A large debate remains as to whether those cells are genuinely neuronal or merely neuronal marker-expressing cells (neuron-like cells), or whether they can effectively integrate into the host brain neuronal connectivity after their graft in vivo
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