Abstract
Understanding how Mesenchymal Stem Cells (MSCs) form blood vessels is critical for creating mechanism-based approaches for the therapeutic use of these cells. In addition, understanding the determinants and factors involved in lineage hierarchy is fundamental to creating accurate and reliable techniques for the study of stem cells in tissue engineering and repair. Dental Pulp Stem Cells (DPSC) from permanent teeth and Stem cells from Human Exfoliated Deciduous teeth (SHED) are particularly interesting sources for tissue engineering as they are easily accessible and expandable. Previously, we have shown that DPSCs and SHEDs can differentiate into endothelial cells and form functional blood vessels through vasculogenesis. Here, we described how we created the “pulpbow” (pulp + rainbow), a multicolor tag experimental model that is stable, permanent, unique to each cell and passed through generations. We used the pulpbow to understand how dental pulp stem cells contributed to blood vessel formation in 3D models in in vitro and ex vivo live cell tracking, and in vivo transplantation assays. Simultaneous tracking of cells during sprout formation revealed that no single multicolor-tagged cell was more prone to vasculogenesis. During this process, there was intense cell motility with minimal proliferation in early time points. In later stages, when the availability of undifferentiated cells around the forming sprout decreased, there was local clonal proliferation mediated by proximity. These results unveiled that the vasculogenesis process mediated by dental pulp stem cells is dynamic and proximity to the sprouting area is critical for cell fate decisions.
Highlights
Postnatal mesenchymal stem cell populations have been found in most healthy and non-healthy oral tissues
Among the most studied dental stem cell populations are the ones derived from the dental pulp of both permanent (DPSCs) and deciduous teeth (SHEDs)
Prior to the creation of the pulpbow, Dental Pulp Stem Cells (DPSC) and Stem cells from Human Exfoliated Deciduous teeth (SHED) were cultured in growth medium composed of α-minimum essential medium (α-MEM; Invitrogen; Waltham, MA, USA) supplemented with 20% fetal bovine serum (FBS; Invitrogen), 1% antibioticantimycotic (Invitrogen) and 20 μg/mL Plasmocin (InVivogen; San Diego, CA, USA)
Summary
Postnatal mesenchymal stem cell populations have been found in most healthy and non-healthy oral tissues. These populations are accessible and can be effortlessly isolated from routinely extracted premolars and 3rd molars or from exfoliating deciduous teeth [1,2] Both SHEDs and/or DPSCs can be used to treat dental and non-dental conditions in preclinical studies [3,4,5] and to have higher proliferative rates [1] and higher regenerative capacity [6] than mesenchymal stem cells (MSCs) derived from the bone marrow. The absence of a vascular network capable of distributing nutrients and oxygen is a major limiting factor [11] In this regard, dental pulp stem cells (SHEDs and DPSCs) can differentiate into vasculogenic endothelial cells both in vitro and in vivo [12,13,14], anastomize with the host vasculature [15] and have higher angiogenic activity than bone marrow MSCs [5]. The precise cellular and molecular mechanisms by which dental pulp stem cells form functional blood vessels remain mostly unclear
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