Formation of 3D stable microvascular networks in a microfluidic chip by supplementing 6-Aminocaproic acid (poster)

Abstract

The formation long-lasting microvascular networks in vitro requires the effective strategies for preventing hydrogel from degradation, in addition to forming stable vasculature. In order to create microvascular network, we cultured human umbilical vein endothelial cells (HUVECs) inside the fibrin gel in microfluidic chips. The cell media applied to the system were supplemented with 6-Aminocaproic acid (EACA) to prevent the fibrin gel from degradation along with cytokines such as vascular endothelial growth factor (VEGF) and angiopoietin 1 (ANG-1) to promote the development of vessels through vasculogenesis. EACA, a potent inhibitor for plasmin, which is the enzyme responsible for fibrinolysis, has been added to the system to minimize the fibrin degradation [1]. To identify complex effects of EACA on vasculogenesis, two different concentrations of EACA (1, 2.5 mg/mL) were tested and compared with control group. Additionally, to optimize proper time points for the addition of EACA into microfluidic devices, the cases of 5 days after seeding HUVECs in devices was tested. For each case, the functionality of microvessels was confirmed by perfusion of microbeads through the vascular networks and staining for endothelial junctions. Our technique for generating stable microvascular networks in microfluidic chips by adding EACA could be further developed to be utilized as a platform for studying long-term vascular diseases such as atherosclerosis or impaired immune system.