The development of methods to isolate and generate human stem cells, along with technology to selectively differentiate them into specific cells and tissue types, has excited many with the promise of improved human cell function understanding, ultimately utilising the cells for regeneration in disease and trauma. However, the two-dimensional growth of derived neurones, which do not mimic the in vivo three-dimensional interactions nor the myriad developmental cues present in vivo, presents itself as a major confounding factor.
Current strategies at producing 3D cultures have focused on using gels or naturally occurring biological molecules such as collagen4 to provide a 3D structure. The nature of these materials is such that their architecture and points of interaction with seeded neurons cannot be predicted or controlled. This leads to essentially random networks with the added disadvantage that the activity of individual elements cannot be readily monitored nor controlled. We aim to revolutionise current neuronal culturing approaches by proposing a novel, highly ambitious interdisciplinary enterprise to construct truly 3D networks that display in vivo activity patterns. This ambition frames the foundational nature of our project. The “MESO-BRAIN” consortium will take the field beyond the state of the art by developing technology and methodologies to produce a new type of neural culture and interacting interface system. The integration of conductive polymers will enable electrical stimulation and recording of individual cells, and the optical properties of the developed scaffold will enable fluorescence imaging and interrogation with photonic/optical approaches. Our project combines new revolutionary tools for micro-fabrication, neuronal network development and monitoring, and functional analysis to bring to light 3D human neuronal networks with tailored characteristics. In naturally developing circuits in the brain, neurons and connections are first configured in a general manner, then cells and networks are gradually refined over time in response to chemical and electrical activity. In our 3D scaffolds, stem cell derived neurons and astrocytes will develop at specific cytophilic points and chemotactic/chemical messages along with electrical activity will be used to promote and drive functional network development. State of the art light sheet imaging platforms will be used to monitor network development and activity.
Functional connectivity maps will be drawn using newly developed and emergent mathematical formulations to verify functional network structure. The foundational nature of our proposal incorporates flexibility in network design, stimulation repertoire and detailed analysis, together with the use of human cells, leading to the reproducible generation of functional 3D human neuronal networks. The provision of human 3D neuronal networks exhibiting physiologically relevant and most importantly reproducible architecture and activity to the science community will be foundational, for the first time enabling large scale scientific investigation of human brain network function, and large scale pharmaceutical testing on human cells and human disease models derived from patients. Availability of 3D human networks will also lead to advances in stem cell derived neural transplantation for CNS therapy and repair.
The Project is made up of six partners from three countries, including:
- Aston University (the Coordinator),
- Axol Bioscience Ltd and Kite Innovation (Europe) Ltd in the United Kingdom
- ICFO- The Institute of Photonic Sciences
- Universitat de Barcelona
- LZH Laser Zentrum Hannover E.V. in Germany.