Human subcutaneous SVF-derived endothelial cells formed venous and arterial capillaries expressing venous markers NR2F2 and CD62E and arterial marker NRP1, with absence of lymphatic endothelial cells in both 2D and 3D cultures.
Human subcutaneous SVF provides an abundant source of venous and arterial blood endothelial cells capable of forming capillaries in 3D models, making it suitable for tissue engineering.
Abstract BACKGROUND: The Stromal Vascular Fraction (SVF) derived from human subcutaneous fat has attracted pivotal interest in clinical applications for its regenerative and anti-inflammatory properties. A deeper characterisation of the endothelial cells within the SVF, across both traditional and tissue-engineered culture systems, is essential for advancing our understanding of endothelial cell biology and enhancing regenerative medicine therapies, including skin substitutes. METHODS: This study investigates endothelial cells from the SVF of human subcutaneous white adipose tissue in 2D culture and 3D bioengineered skin models to better define their specific subtypes. Immunofluorescence analysis was used to assess the SVF, with particular focus on endothelial cells, including their ability to form capillary-like networks within type I collagen hydrogels. RESULTS: Analysis of the SVF-derived cells showed PLVAP-positive blood endothelial cells but no lymphatic endothelial cells. The blood endothelial cells could be discriminated into NR2F2- and CD62E-positive venous endothelial cells and NRP1-expressing arterial endothelial cells. Within the 3D hydrogels, the blood endothelial cells formed venous and arterial capillaries. CONCLUSION: We characterised endothelial cells from human subcutaneous SVF, identifying venous and arterial blood endothelial cells while confirming the absence of lymphatic endothelial cells in vitro . These findings underline that subcutaneous adipose tissue is an attractive cell source due to its ease of isolation and abundance of endothelial cells for skin tissue engineering and regenerative medicine in general.
Schwager et al. (Wed,) conducted a other in Adult healthy individuals undergoing subcutaneous fat liposuction and juvenile humans with foreskin tissue for endothelial cell characterization. Human subcutaneous stromal vascular fraction derived endothelial cells (SVF-derived ECs) cultured in 2D and 3D systems vs. Human dermal microvascular endothelial cells (HDMECs) cultured in 2D and 3D was evaluated on Identification and characterization of endothelial cell subtypes (arterial, venous, lymphatic) via immunofluorescence markers in 2D culture and 3D collagen type I hydrogels. Human subcutaneous SVF-derived endothelial cells formed venous and arterial capillaries expressing venous markers NR2F2 and CD62E and arterial marker NRP1, with absence of lymphatic endothelial cells in both 2D and 3D cultures.
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