Thesis Project Form

To better mimic the complex connectivity found in the human brain, many efforts have been devoted to build controllable in vitro neuronal structures with a 3D architecture. The main goal of this thesis is to build and characterize both 2D and 3D networks in vitro composed by iNeurons, differentiated from human-induced pluripotent stem cells (h-iPSCs), in combination with cortical rat astrocytes. The 3D structure will be guaranteed by the use of a thermosensitive biopolymeric hydrogel, where cells can be encapsulated.

The spontaneous activity of both 2D and 3D networks will be acquired at different time points during the development by means of planar MEAs. The possibility to use an acquisition system based on probes, typically used for in vivo recordings, which can penetrate the 3D tissue will be evaluated and explored.
The main activities of the thesis include, but may not be limited to:
- In vitro experiments. Experiments with 2D and 3D in vitro cultures of neurons with microelectrode arrays acquisitions and, possibly, probes. The assignee will be actively involved in all the experimental tasks, in order to perform the experiments in autonomy.
- Data processing. Neuronal data will be processed using the tools available in the Lab. Further algorithms for the analysis will be performed and implemented by the assignee.
- Comparative analysis and data interpretation. Data from 2D and 3D cultures will be compared and a developmental profile for each experimental group provided.
The knowledge gained during this project will help to understand the electrophysiological phenotypes of 2D and 3D cultures and will allow to make a step forward towards the development of ‘brain-like’ models to be exploited for patient-specific therapeutics.

1. Experimental skills: electrophysiology, imaging, biopolymeric hydrogels preparation and characterization
2. Mixed software skills (e.g. MATLAB)
3. Neurophysiological data analysis and interpretation