Human dermal microvascular arterial and venous blood endothelial cells and their use in bioengineered dermo-epidermal skin substitutes in vitro and in vivo

<p>The bio-engineering of vascular networks is pivotal to create complex tissues and</p> <p>organs in vitro for regenerative medicine applications. The vascular plexus is needed for a</p> <p>sufficient and fast blood supply after transplantation, and, thus, required for the survival and</p> <p>function of the engineered tissue or organ. Hence, human endothelial cells are an attractive</p> <p>source for bio-engineering purposes, for example human dermal microvascular endothelial</p> <p>cells (HDMECs).</p> <p>So far, a discrimination between arterial and venous blood endothelial cells after</p> <p>isolation of HDMECs from skin biopsies and if arterial and/or venous capillaries are formed in</p> <p>pre-vascularized bio-engineered substitutes was not investigated.</p> <p>In this study, we investigated employedby single cell sequencing for to</p> <p>investigate/compare human arterial and venous endothelial cell markers in human fetal and</p> <p>juvenile skin. Further, we analyzed if these markers are present after isolation of human skin</p> <p>derived endothelial cells under 2D culture conditions. In additionFinally, we investigated</p> <p>assessed if human endothelial cells form distinct arterial and venous capillaries in 3D</p> <p>collagen type I hydrogels, and if these capillaries retain their identity after transplantation.</p> <p>We determinedOur results showed that arterial and venous endothelial cell markers</p> <p>such as NRP1 and NR2F2 are expressed both in fetal and juvenile skin, and are retained after</p> <p>isolation in culture. We could show demonstrate that arterial and venous endothelial cells</p> <p>maintain their differentiation status and form arterial and venous capillaries in 3D in vitro</p> <p>culture systems and that the capillaries inosculate after transplantation.</p> <p>In summary, we could show that we could bio-engineer human arterial, venous, and</p> <p>lymphatic capillaries in a human skin substitute in view of regenerative medicine approaches</p> <p>for clinical applications.</p>