Project No. 2479

STANDARD PROJECT

Primary Supervisor

Dr Sandrine Willaime-Morawek – University of Southampton

Co-Supervisor(s)

Dr Andrew Penn – University of Sussex

Prof Sumeet Mahajan – University of Southampton

Prof Amrit Mudher – University of Southampton

Summary

3D culture models provide a physiologically and spatially relevant microenvironment thereby aiding better differentiation and maturation of cells in vitro.

We developed a 3D neural model derived from healthy and Alzheimer’s disease patients iPSCs, in which cells differentiate, self-organize and mature. iPSCs were differentiated in 3D Matrigel for 18 weeks in vitro. The 3D model was characterized for cell morphology and protein profile. iPSCs differentiate into neurons and astrocytes which self-organize into 3D structures by 3 weeks of differentiation in vitro. Cells express astrocytic, neuronal, glutamatergic, GABAergic and pre-synaptic markers after differentiation.

To improve this model, it needs validation against known maturity and ageing benchmarks, by exploring protein biochemistry (mature isoforms of proteins and ageing protein aggregation), as well as optimisation with live label-free 3D imaging and access for concomitant electrophysiology to validate neuronal maturity and functionality.

The overall aim of this interdisciplinary PhD project is thus to develop & validate both our existing 3D neural cell culture model and a novel imaging methodology to study the biochemistry of ageing.

The project is divided into 3 aims:

1. Improve the 3D model setup to allow concomitant imaging and electrophysiology.

2. Optimise light-sheet imaging-based methodology to study protein biochemistry in 3D neural cultures and validate it against known ageing markers, including using techniques such as Atomic Force microscopy (AFM), Raman Spectroscopy and Dynamic Light Scattering (DLS).

3. Validate maturity and ageing of the 3D neuronal network using electrophysiology techniques.

The project can potentially provide unprecedented insight into ageing biochemistry. This will allow better in vitro modelling of complex neurological diseases and result in a novel methodology for studying 3D cultures. Impacts also include training of future researchers in cutting-edge techniques and methodologies.

For this project, a self-driven candidate motivated by interdisciplinary collaboration, with innovative thinking and adaptability, would be most suitable.