Paving the way for High-throughput Organoid ENgineering using Integrated acoustiX

Introduction

The aim of PHOENIX is to use my expertise in microsystems engineering to close critical technology gaps in organoid generation. Cerebral organoids are 3D self-assembled structures derived from human induced pluripotent stem cells, replicating both structure and function of the human foetal brain. Organoids have the potential to replace existing 2D cell cultures and animal models, but this has not yet been realised due to rudimentary preparation methods.

Technology Gaps

In PHOENIX, three important technology gaps will be addressed:

1. Reproducibility
2. Controlled maturation
3. Vascularisation

I aim to build on my pioneering research on droplet acoustofluidics and the scientific output of my ERC Starting Grant to develop three microfluidic modules that, at the end of the project, shall be integrated into a seamless organoid engineering pipeline.

Methodology

The technology in focus is acoustophoresis, a method to manipulate particles and cells by ultrasound. This will be used to achieve ordered encapsulation of stem cells in hydrogel droplets and develop a microfluidic platform where the cells can be differentiated under fully controlled conditions.

Vascular Network Integration

Finally, two-photon writing will be used to integrate a vascular network with the organoid constructs to form an important delivery architecture for nutrients and blood components. PHOENIX will be focused on both technology development and thorough biological characterisation of the resulting organoids to demonstrate both expected and unexpected benefits of transferring organoid generation on-chip.

Collaborations

Collaborations have been established with:

• Prof. Christine Mummery at LUMC, NL
• Dr. Valeria Orlova at LUMC, NL
• Dr. Anna Falk at KI, SE

These collaborations will provide expertise in complementary fields of this highly interdisciplinary project.

Expected Output

The expected output of PHOENIX is a microfluidic technology that enables high-throughput generation of cerebral organoids with a multi-regional structure and vascularisation in a direct process.