Our angiogenesis assay was created during the project “Growing real human small blood vessels in the laboratorium”, in which Ncardia participated together with with Leiden University Medical Center, Leiden Academic Center for Drug Research and Mimetas. The aim of this project was to develop a next generation capillary-on-a-chip model to study mechanisms underlying small blood vessel loss in disease such as ‘heart failure with preserved ejection fraction’.

In the angiogenisis assay, hiPSC-derived endothelial cells are grown in a microfluidic channel. Upon triggering with a gradient of proangiogenic factors, the sprouting cells form an intricate capillary network in an adjacent 3-dimensional matrix. The angiogenesis assay platform (48 devices per plate) is compatible with automated systems for cell plating and imaging, to enable secondary screening.

Click here to watch how our angiogenesis assay was created: Standardized and scalable angiogenesis assay

Inhibitory effects of anti-angiogenic compounds

This case study is part of the project “Growing real human small blood vessels in the laboratorium”.

We developed a perfused 3D angiogenesis assay that includes endothelial cells (ECs) from induced pluripotent stem cells (iPSC) and assessed its performance and suitability for anti-angiogenic drug screening. Angiogenic sprouting was compared with primary ECs and showed that the microvessels from iPSC-EC exhibit similar sprouting behavior, including tip cell formation, directional sprouting and lumen formation.

In this project, we optimized the concentration for sunitinib inhibition, and showed that sunitinib inhibits sprouting from concentrations >10 nM, while angiogenesis is completely inhibited at concentrations >50 nM. Angiogenic sprouts of IPSC-EC's are shown after 48 hours with various concentrations of Sunitinib. (Yellow = F-actin, blue = nucleus). The graphs below show a quantitative analysis, demonstrating the effect of 50 nM Sunitinib.

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This data was obtained in collaboration with Mimetas and LUMC.

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