Vascularization is one of the most central processes enabling multicellular life. Owing to the complexity of vascularization regulatory networks, minor control imbalances often have severe pathologic consequences ranging from ischemic diseases to cancer. Tissue engineers are immediately confronted with vascularization as artificial tissues of a clinically relevant size require a vascular system to ensure vital physiologic logistics throughout the entire tissue and to enable rapid connection to the host vasculature following implantation. Using human umbilical vein endothelial cells (HUVECs) coated onto a human aortic fibroblast (HAF) core microtissue generated by gravity-enforced self-assembly in hanging drops, we created a tissue-culture system for studying capillary network formation. We provide comprehensive technical insight into the design and analysis of prototype vascularization in multicell-type-based microtissues. Detailed understanding of generic processes managing capillary formation in human tissue culture may foster advances in the development of clinical tissue implants.
