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

The bioengineering of vascular networks is pivotal to creating complex
tissues and organs for regenerative medicine applications. However,
bioengineered tissues comprising an arterial and venous plexus alongside a
lymphatic capillary network have not been explored yet. Here, we first
employed scRNA-seq to investigate the arterio-venous endothelial cell
marker patterning in human fetal and juvenile skin. Transcriptomic
analysis revealed that arterial and venous endothelial cell markers NRP1
and NR2F2 are broadly expressed in fetal and juvenile skin. In contrast,
expression of NRP1 and NR2F2 on the protein level was cell-type specific
and was retained in 2D cultures in vitro. Finally, we bioengineered
distinct arterial and venous capillaries in 3D hydrogels and demonstrated
rapid anastomosis with the host vasculature in vivo. In summary, we
established the bioengineering of human arterial, venous, and lymphatic
capillaries, hence paving the way for these cells to be used in
regenerative medicine and future clinical applications.