A well plate-based multiplexed platform for incorporation of organoids into an organ-on-a-chip system with a perfusable vasculature

Owing to their high spatiotemporal precision and adaptability to different host cells, organ-on-a-chip systems are showing great promise in drug discovery, developmental biology studies and disease modeling. However, many current micro-engineered biomimetic systems are limited in technological application because of culture media mixing that does not allow direct incorporation of techniques from stem cell biology, such as organoids. Here, we describe a detailed alternative method to cultivate millimeter-scale functional vascularized tissues on a biofabricated platform, termed 'integrated vasculature for assessing dynamic events', that enables facile incorporation of organoid technology. Utilizing the 3D stamping technique with a synthetic polymeric elastomer, a scaffold termed 'AngioTube' is generated with a central microchannel that has the mechanical stability to support a perfusable vascular system and the self-assembly of various parenchymal tissues. We demonstrate an increase in user familiarity and content analysis by situating the scaffold on a footprint of a 96-well plate. Uniquely, the platform can be used for facile connection of two or more tissue compartments in series through a common vasculature. Built-in micropores enable the studies of cell invasion involved in both angiogenesis and metastasis. We describe how this protocol can be applied to create both vascularized cardiac and hepatic tissues, metastatic breast cancer tissue and personalized pancreatic cancer tissue through incorporation of patient-derived organoids. Platform assembly to populating the scaffold with cells of interest into perfusable functional vascularized tissue will require 12-14 d and an additional 4 d if pre-polymer and master molds are needed. This protocol describes how to integrate organoids into a vascularized organ-on-a-chip biofabricated platform. The platform is assembled in a 96-well format and allows the connection of two tissues through a single endothelialized microchannel blood vessel.

Automated summary

The study presents an alternative method for cultivating millimeter-scale functional vascularized tissues on a biofabricated platform, facilitating the easy integration of organoid technology. By utilizing 3D stamping with a synthetic polymeric elastomer, a scaffold called 'AngioTube' is generated to support a perfusable vascular system and the self-assembly of various parenchymal tissues. The platform allows easy connection of two or more tissue compartments through a common vasculature, with built-in micropores for studying cell invasion.