Vascularization of multi-Organ-on-Chips with blood and lymphatic endothelial cells for the generation of immunocompetent skin models

Description

Background 3D human skin models fail to include both blood (BEC) and lymphatic (LEC) endothelial cells despite their essential role for homeostasis and immune responses, limiting their relevance for disease modeling and safety testing.
Aim Establish a method for vascularization of organ-on-chip microfluidics with human BECs or LECs which allows long term culturing under physiologic flow conditions for immune competent (multi-) organ-on-chip models.
Methods Primary human skin endothelial cells were separated into BECS and LECs, expanded to generate, used to vascularize multi-organ-on-chip microfluidic bioreactors and cultured for 14 days under dynamic flow mimicking blood and lymph flow pressures.
Results Large numbers of highly pure BECS and LECs were obtained and used to vascularize organ-on-chip devices. Upon prolonged culture, cells retained their endothelial specific phenotype. Mimicking blood vessel flow induced morphological changes as cells aligned in the direction of the flow, while this does not occur when applying lower lymphatic vessel flow. BEC and LEC biomarker expression of was clearly different but not influenced by flow conditions. BECs and LECs respond to inflammatory conditions by upregulating soluble ICAM, VCAM, CCL2 and IL-6. While mRNA levels of endothelial junction markers (Cldn5, VEcadh, ZO1) did not change in BECs and LECs, the LEC specific marker Prox-1 was clearly reduced in LECs upon inflammatory conditions.
Conclusion: The presented method can be used to further enhance organ-on-chip models through the incorporation of functional blood and lymphatic endothelial cells. This will result in relevant healthy and diseased tissue models to investigate human disease and safety testing.

Period	27 Aug 2023 → 31 Aug 2023
Event title	12th World Congress on Alternatives and Animal Use in the Life Sciences (WC12)
Event type	Conference
Location	Niagra, CanadaShow on map
Degree of Recognition	International