HUPERVAS-CHIP: minimal essentials for humanized vessel-on-a-chip model for vascular research

Abstract Introduction and Objectives: Three-dimensional in vitro organ-on-a-chip models already represent a significant advance in biotechnology research, providing effective, predictive, and cost-efficient preclinical models that more closely recapitulate tissue physiology. Our objective was to develop a humanized personalized vessel-on-a-chip model (HUPERVAS-CHIP) using a biocompatible scaffold that supports cell growth and bilayer coculture of human vascular smooth muscle cells (VSMC) and human umbilical vein endothelial cells (HUVEC), with the capacity to be integrated into microfluidic systems. In the long term, we aim is to employ this model as part of a more personalized and precise medicine approach. Methods VSMCs and HUVECs were isolated from donated umbilical cords following informed consent. After characterization by flow cytometry and immunofluorescence, cells were cultured in polydimethylsiloxane (PDMS) chips containing channels with an internal diameter of 1 mm. Cell-specific culture media (SmGM-2 and EGM-2), supplemented with tropoelastin, were used to promote activation and extracellular matrix (ECM) formation, as well as to facilitate HUVEC growth over the VSMC layer. Then, using a microfluidic pump system, the functional relevance of the model was evaluated by fluidic and permeability studies after 9 days-term cell-growing period. Results Optimal isolation and culture conditions were established to enable the formation of a vascular unit composed of VSMCs and HUVECs within the vascular chip. After nine days of coculture, the model exhibited the main components of the ECM. Functional studies demonstrated that a flow rate of 1 mL/min enhanced cell adhesion and endothelial monolayer integrity more effectively than static conditions or lower flow rates. Permeability assays using dextran molecules of defined molecular weight provided physiological validation of the model. Conclusions HUPERVAS-CHIP has demonstrated its potential as a robust preclinical research model that preserves the essential structural and functional features of blood vessels, incorporates their principal effector cells, and recapitulates key physiological parameters, including cellular maturation, shear stress responsiveness, and functional permeability. Accordingly, it represents a promising platform for the evaluation of vasoactive compounds and the assessment of vascular toxicity. Moreover, it offers a valuable tool for elucidating the molecular mechanisms underlying cardiovascular diseases.