United Kingdom
A 9-week laboratory-based research project under the supervision of Dr. Michael Lattke in the Guillemot Lab at the Francis Crick Institute, as part of the Crick Calleva Summer Student Program. Homogenous populations of mature astrocytes can be used to investigate neurodegenerative disorders in vitro. However, current in vitro differentiation protocols generate mostly immature astrocytes, and the lack of markers to characterise mature astrocytes makes it difficult to optimise culturing conditions. My project aims to improve the in vitro differentiation protocol of astrocytes from murine neural stem cells. To this end, murine neural stem cells from three different cell lines were cultured in the presence of different combination of supplements for 14 days. qPCR was used to analyse the RNA expression, and immunostaining was used to analyse the protein expression of a set of astrocytic maturation markers identified through RNA-seq of mature astrocytes in vivo. I was able to show that 2D tissue culturing conditions only upregulated a limited set of the astrocytic maturation markers at the RNA and protein level, whereas culturing murine neural stem cells in 3D as a neurosphere improves the percentage of astrocytic maturation marker upregulated. This project allowed me to learn RNA extraction, qPCR, and immunohistochemistry techniques, and I was also able to reinforce my skills in tissue-culture and confocal microscopy.
Elected Co-President of Churchill College Medical & Veterinary Society, University of Cambridge, for the 2017/18 academic year. The responsibilities include organising academic, welfare and social events for all medical and veterinary students at Churchill College. For example, a peer mentoring scheme, the annual MedSoc dinner, and informative talks throughout the year.
An 8-week laboratory-based research project under the supervision of Dr. Lewis Evans in the Livesey Lab at the Gurdon Institute. Hernandez-Vega et al. (2017) have shown the ability of tau protein to form liquid-like droplets in vitro (phase transition), and the accumulation of tubulin protein in these tau droplets to form microtubules. My project aims to verify these finding using wildtype 2N4R tau and investigate whether P301S mutant tau has different properties in phase transition and microtubule nucleation. Tau protein promotes microtubule formation giving the intracellular neurofibrillary tangles seen in patients with Alzheimer’s disease, the P301S mutant is highly aggregation prone and has been linked with frontotemporal dementia. After successfully cloning, expressing and purifying WT and P301S mutant tau proteins in E. Coli, I was able to show the liquid-liquid phase transition of tau proteins in vitro, and the subsequent polymerisation of microtubules when tubulin and GTP were added to the tau “droplets”. Overall, I was able to develop an assay which could be used to investigate the differences between wildtype and mutant forms of tau on microtubule nucleation and stabilization. This project has given me the opportunity to learn DNA cloning, bacterial transformation, DNA gel extraction, protein purification and confocal microscopy techniques. I am also able build on my previous experiences in western blotting and tissue-culture, in particular I had the opportunity to work with human iPS cell-lines.