As engineered tissues progress toward therapeutically relevant length scales and cell densities, it is critical to deliver oxygen and nutrients throughout the tissue volume via perfusion through vascular networks. Furthermore, seeding of endothelial cells within these networks can recapitulate the barrier function and vascular physiology of native blood vessels. In this protocol, we describe how to fabricate and assemble customizable open-source tissue perfusion chambers and catheterize tissue constructs inside them. Human endothelial cells are seeded along the lumenal surfaces of the tissue constructs, which are subsequently connected to fluid pumping equipment. The protocol is agnostic with respect to biofabrication methodology as well as cell and material composition, and thus can enable a wide variety of experimental designs. It takes ~14 h over the course of 3 d to prepare perfusion chambers and begin a perfusion experiment. We envision that this protocol will facilitate the adoption and standardization of perfusion tissue culture methods across the fields of biomaterials and tissue engineering.

随着工程组织朝着治疗相关的长度尺度和细胞密度发展，通过血管网络灌注在整个组织体积内输送氧气和营养物质至关重要。此外，在这些网络中植入内皮细胞可以重现天然血管的屏障功能和血管生理学。在本协议中，我们描述了如何制造和组装可定制的开源组织灌注室，以及如何在其中插入组织结构。人内皮细胞沿着组织结构的腔表面接种，随后连接到流体泵送设备。该协议在生物制造方法以及细胞和材料成分方面是不可知的，因此可以实现各种各样的实验设计。在 3 天的过程中需要 ~ 14 小时来准备灌注室并开始灌注实验。我们设想，该协议将促进生物材料和组织工程领域灌注组织培养方法的采用和标准化。