An Arteriovenous Bioreactor Perfusion System for Physiological In Vitro Culture of Complex Vascularized Tissue Constructs.

backgroundThe generation and perfusion of complex vascularized tissues in vitro requires sophisticated perfusion techniques. For multiscale arteriovenous networks, not only the arterial, but also the venous, biomechanical and biochemical conditions that physiologically exist in the human body must be accurately emulated. For this, we here present a modular arteriovenous perfusion system for the in vitro culture of a multi-scale bioartificial vascular network.

methodsThe custom-built perfusion system consisted of two circuits: in the arterial circuit, physiological arterial biomechanical and biochemical conditions were simulated using a modular set-up with a pulsatile peristaltic pump, compliance chambers, and resistors. In the venous circuit, venous conditions were emulated accordingly. In the center of the system, a bioartificial multi-scale vascularized fibrin-based tissue was perfused by both circuits simultaneously under biomimetic arteriovenous conditions. Culture conditions were monitored continuously using a multi-sensor monitoring system.

resultsThe physiological arterial and venous pressure- and flow-curves, as well as the microvascular arteriovenous oxygen partial pressure gradient, were accurately emulated in the perfusion system. The multi-sensor monitoring system facilitated live monitoring of the respective parameters and data-logging. In a proof-of-concept experiment, vascularized three-dimensional fibrin tissues showed sustained cell viability and homogenous microvessel formation after culture in the perfusion system.

conclusionsThe arteriovenous perfusion system facilitated the in vitro culture of a multiscale vascularized tissue under physiological pressure-, flow-, and oxygen-gradient conditions. With that, it presents a promising technique for the in vitro generation and culture of complex large-scale vascularized tissues.