VEGFR1 primes a unique cohort of dental pulp stem cells for vasculogenic differentiation

Dental Pulp Stem Cells (DPSC) constitute a unique group of cells endowed with multipotency, self-renewal, and the capacity to regenerate the dental pulp tissue. While much has been learned about these cells in recent years, it is still unclear if each DPSC cell is multipotent or if unique sub-populations of DPSCs are “primed” to undergo specific differentiation paths. The purpose of this study was to define whether a sub-population of DPSCs is uniquely primed to undergo vasculogenic differentiation. Here, permanent tooth DPSCs or Stem cells from Human Exfoliated Deciduous teeth (SHED) were flow sorted for VEGFR1 and exposed to vasculogenic differentiation medium, i.e. Endothelial Growth Medium (EGM) 2-MV supplemented with 50 ng/ml rhVEGF(165) in presence of 0 or 25 μg/ml anti-human VEGF antibody (bevacizumab; Genentech). In addition, sorted SHED (i.e. VEGFR1(high) or VEGFR1(low)) were seeded in biodegradable scaffolds and transplanted into the subcutaneous space of immunodeficient mice. Despite proliferating at a similar rate, VEGFR1(high) generated more in vitro sprouts than VEGFR1(low) cells (p<0.05). Blockade of VEGF signaling with bevacizumab inhibited VEGFR1(high)-derived sprouts, demonstrating specificity of responses. Similarly, VEGFR1(high) SHED generated more blood vessels when transplanted into murine hosts than VEGFR1(low) cells (p<0.05). Collectively, these data demonstrate that dental pulp stem cells contain a unique sub-population of cells defined by high VEGFR1 expression that are primed to differentiate into vascular endothelial cells. These data raise the possibility of purifying stem cells with high vasculogenic potential for regeneration of vascularized tissues or for vascular engineering in the treatment of ischemic conditions.