On‐Chip Reconstitution of Uniformly Shear‐Sensing 3D Matrix‐Embedded Multicellular Blood Microvessel

Preclinical human‐relevant modeling of organ‐specific vasculature offers a unique opportunity to recreate pathophysiological intercellular, tissue‐tissue, and cell‐matrix interactions for a broad range of applications. Here, this work presents a reliable, and simply reproducible process for constructing user‐controlled long rounded extracellular matrix (ECM) embedded vascular microlumens on‐chip for endothelization and co‐culture with stromal cells obtained from human lung. This work demonstrates the critical impact of microchannel cross‐sectional geometry and length on uniform distribution and magnitude of vascular wall shear stress, which is key when emulating in vivo observed blood flow biomechanics in health and disease. In addition, this study provides an optimization protocol for multicellular culture and functional validation of the system. Moreover, this study shows the ability to finely tune rheology of the three‐dimensional natural matrix surrounding the vascular microchannel to match pathophysiological stiffness. In summary, this work provides the scientific community with a matrix‐embedded microvasculature on‐chip populated with all‐primary human‐derived pulmonary endothelial cells and fibroblasts to recapitulate and interrogate lung parenchymal biology, physiological responses, vascular biomechanics, and disease biogenesis in vitro. Such a mix‐and‐match synthetic platform can be feasibly adapted to study blood vessels, matrix, and ECM‐embedded cells in other organs and be cellularized with additional stromal cells. This study presents design, development, and validation of a reliable and highly reproducible process for constructing user‐controlled long rounded extracellular matrix (ECM) embedded vascular microlumens on‐chip for endothelization and co‐culture with stromal cells. Organ‐specific (lung in this case) primary human‐derived endothelial cells, fibroblasts, and stiffness‐adjustable hydrogels are used to recreate microvascular (MV) tissue architecture, physiological biology, vascular biomechanics, and disease biogenesis in vitro. The method provides a state‐of‐the‐art microphysiological system (MPS, referred to as “ECM MV‐Chip”) that allows concurrent analysis of blood microvessels, perivascular matrix, and ECM‐embedded stromal cells.