Flow in fetoplacental microvessels in vitro enhances perfusion, barrier function, and matrix stability

Abstract Proper placental vascularization is vital for pregnancy outcomes, but assessing it with animal models and human explants has limitations. Here, we present a 3D in vitro model of human placenta terminal villi that includes fetal mesenchyme and vascular endothelium. By co-culturing HUVEC, placental fibroblasts, and pericytes in a macro-fluidic chip with a flow reservoir, we generate fully perfusable fetal microvessels. Pressure-driven flow is crucial for the growth and remodeling of these microvessels, resulting in early formation of interconnected placental vascular networks and maintained viability. Computational fluid dynamics simulations predict shear forces, which increase microtissue stiffness, decrease diffusivity and enhance barrier function as shear stress rises. Mass-spec analysis reveals the deposition of numerous extracellular proteins, with flow notably enhancing the expression of matrix stability regulators, proteins associated with actin dynamics, and cytoskeleton organization. Our model provides a powerful tool for deducing complex in vivo parameters, such as shear stress on developing vascularized placental tissue, and holds promise for unraveling gestational disorders related to the vasculature.