A fluid-walled microfluidic platform for human neuronal microcircuits and axotomy
Authors/Creators
- 1. Oxford Parkinson's Disease Centre and Department of Physiology, Anatomy and Genetics, University of Oxford, South Park Road, Oxford OX1 3QU, United Kingdom; Kavli Institute for Neuroscience Discovery, University of Oxford, Dorothy Crowfoot Hodgkin Building, South Park Road, Oxford OX1 3QU, United Kingdom; Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815, USA
- 2. Osney Thermofluids Institute, Department of Engineering Science, University of Oxford, Osney Mead, Oxford OX2 0ES, United Kingdom; The Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom.
- 3. The Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom.
- 4. Osney Thermofluids Institute, Department of Engineering Science, University of Oxford, Osney Mead, Oxford OX2 0ES, United Kingdom
Description
ABSTRACT
Recapitulation of human neuronal circuits in vitro will significantly facilitate investigation of human neuronal physiology and pathophysiology. This paper demonstrates and characterises a novel system for culturing neuronal circuits within fluid-walled microenvironments. Fluid-walled technology leverages the immiscibility between aqueous cell media and hydrophobic bio-inert fluorocarbon (FC40) to create miniaturised cell cultures inside standard Petri dishes. Here, we demonstrate the capacity of fluid-walled technology in recreating the human corticostriatal microcircuit. Induced pluripotent stem cell-derived cortical and striatal neurons were co-cultured in fluid-walled dumbbells which, employing intrinsic pressures, allows i) separation of different neuronal subtypes, ii) compartmentalisation of cortical somas and dendrites from cortical axons, and iii) unidirectional projection of cortical axons toward their postsynaptic striatal partner. Exploiting unique features of fluid walls, we also describe a contactless method to perform localised cortical axotomy at pre-defined location along the dumbbells using a micro-jet of cell media. Axotomy of all cortical axonal tracts on the same dish is systematically obtained within 90 seconds. Subsequent axonal regeneration was observed and significantly promoted in the presence of postsynaptic striatal neurons. The ability to control compartmentalisation and directionality of neuronal connections based on fluid dynamics offers an efficient and highly reproducible system for long-term culturing of human neuronal circuits over conventional monoculture of cortical neurons. Combined with reproducible axotomy, we envision this system to be adaptable to large scale screening of regenerative therapeutic candidates in a highly physiological manner.
FILE DESCRIPTIONS
Source Data.xlsx: Tabular datasets plotted on main figures 2 and 5.
Key Resources Table.xlsx: Table containing key resources (primary and secondary antibodies, cell lines and software) used in this study.
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Additional details
Funding
- Aligning Science Across Parkinson's
- Mapping the modulatory landscape governing striatal dopamine signaling and its dysregulation in Parkinson’s disease ASAP-020370