Over the course of the two weeks of my work experience, I had the opportunity to try out new laboratory techniques and to explore various aspects of the research being done at the University of Birmingham, Centre for Liver Research. In this account I will focus solely on the major highlights (despite each day having been amazing and enriching in its own right).
On the first day, I was greeted warmly by Dr Zania Stamataki and given a tour of the Medical School building and the IBR building. We discussed about the research being done by her team concerning hepatocytes and their interaction with the immune system (mainly CD4+ T-cells). Having completed a FutureLearn course on the liver (Liver Disease: Looking after Your Liver – University of Birmingham) provided me with a decent background to understanding the fascinating concepts Zania introduced me to. I learnt the basics of cell-in-cell structures and the vital efferocytosis that hepatocytes perform and what these processes mean for inflammation and the development of cancer and metastasis.
In the afternoon, Janine Fear lead an induction in which I was exposed to the many fascinating laboratories and ‘behind-the-scene’ areas such as the waste disposal and incineration systems, the autoclave room, and the liquid nitrogen cooling systems. One of the many surprising things I was shown was the autoclave tape – I couldn’t help but marvel at the simple yet ingenious concept of tape that changes colour when exposed to the high temperatures of the sterilisation process. Janine Fear also informed me of the lab safety procedures and that the labs I would be working in are classed as Category 2 and that the majority of the labs are fitted with Class II biosafety cabinets. As part of the induction, Janine also taught me basic skills (such as how to use a micropipette and a pipette gun) which I later utilised when performing experiments.
After the induction, I was introduced to Adam McGuinness (a PhD student) who showed me several culture flasks (containing hepatocytes and T-cells) under a light microscope. He also explained that the nutrient medium provided to the cells contains an indicator which changes to yellow as the solution becomes acidic (because CO2 is produced as the cells metabolise) – indicating that the medium should be changed. I recalled that a similar process was used by the BactAlert machines in the serology lab in Mumbai (where I shadowed Dr Salunke last year); the machine detected low pH of blood cultures (which indicates sepsis: bacteria in the blood produce CO2 – making the blood acidic). The hepatocytes cultured in this lab were from a cancer cell line (Huh7) – I learnt that cell lines are ideal for research as they can survive and replicate better than primary cells in vitro.
On day two, Zania and I discussed the role of a researcher and the many facets of the job (including the process of writing and revising publications and some aspects of what makes for a high-impact publication worthy of a renowned journal such as Cell or Nature). Our discussion then drifted back to Zania’s current research and she explained to me that the host cells of cell-in-cell structures can fail to divide as the cell inside them becomes a physical obstruction to cytokinesis. Fascinated by this concept, I spent the rest of the morning reading publications* on cell-in-cell structures which Zania recommended to me (including one written by her former PhD student, Alex Wilkinson).
In the afternoon, I observed Adam performing an experiment to image a host cell hepatocyte failing to divide due to a T-cell inside it. He explained the various fluorescent cell tracker dyes and cytoplasm stains which would be used to image the experiment. We then visited Dr Omar Qureshi who introduced us to the impressive Celldiscoverer 7 (ZEISS) capable of fluorescence microscopy and performing time-lapse imaging.
The next day, imaging experts from ZEISS explained the process of analysing the images produced by the Celldiscoverer 7; the analysis software can be configured to automatically count the number of cell-in-cell structures and identify any host cells which failed to divide.
Afterwards, I followed Adam to a meeting with the rest of the students who are taking the same PhD course (Sci-Phy). One of the major advantages of shadowing both a student and a PhD supervisor was that I gained exposure to the many aspects of completing a PhD course (such as writing a thesis and preparing for a viva) from different perspectives. During this meeting, Adam presented his plans for a mini-project which involved using machine learning to identify and quantify cell-in-cell structures.
In the afternoon, I attended an interesting lecture (presented by Alejandra Tomas from Imperial College) with Zania regarding current research into improving pancreatic beta cell function.
On the fourth day, I had the very exciting chance to do some ‘hands-on’ work; under the direction of Zania, I ‘split’ a cell culture of hepatocytes. Since the hepatocytes are from a cancer cell line, they can be repeatedly ‘split’ to allow them to proliferate and be used for various experiments. It was delightful to have this opportunity which enabled me to sharpen my skills in handling pipettes and working with the hepatocytes. I also learnt how to care for the surfaces and machines in the lab; I cleaned all the worktops in the labs with 70% IMS (Industrial Methylated Spirit) and Trigene before commencing my work, which ensures that the surfaces are sterile.
The afternoon provided another opportunity to immerse myself further in the topic of cell-in-cell structures and I read two absorbing publications: ‘Cell-in-cell phenomena, cannibalism, and autophagy’ and ‘Entosis enables a population response to starvation’. I found the latter article intriguing as it introduced the idea of host cells obtaining nutrients and biomass through phagocytosis of neighbouring cells as part of a coordinated response to long-term glucose starvation.
On day five, I observed Adam perform another experiment which explored the relationship between the number of necrotic T-cells provided to hepatocytes and the number of phagocytosis events (in which cell-in-cell structures are formed). The experiment used serial dilutions of necrotic T-cells; hepatocytes can engulf dead T-cells easily (even in low quantities). Following this, we imaged the experiment using the smaller, yet equally impressive, CX5 fluorescence microscope.
In the afternoon, we analysed the images from this experiment to yield averages for the number of cell-in-cell structures observed with the different concentrations of T-cells. I performed some manual counts with the help of an image editing software (Image J) whilst Adam began to use Matlab (a programming language) to automate this process.
On the first day of my second week with the University of Birmingham, I attended a conference with Zania (who chaired the third session) on stromal microenvironments in health and disease. A variety of speakers from the University of Birmingham (and a plenary speaker from Barts Cancer Institute) presented their research on a vast range of topics connected to stromal cells (cells that make up the supporting tissue of organs) from potential cures for rheumatoid arthritis (presented by Andrew Filer) to identifying indicators of myeloma and hallmarks of cancer (presented by Dan Tenant). These presentations helped to further enhance my understanding of the incredible research undertaken at the University of Birmingham.
Two days later, I had the pleasure of meeting visiting Professor Lucie Peduto from the Pasteur Institute. Her research lab has a strong focus on stroma which relates to the liver stromal microenvironment researched here. In the afternoon, I attended her seminar on the relationship between stromal cells, fibrosis, and cancer; having attended the conference on stromal microenvironments earlier that week allowed me to gain a better understanding of her research.
The penultimate two days of my work experience were by far my favourite; I had the exciting opportunity to perform an entire experiment single-handedly. My detailed observation of the experiments performed by Adam proved very useful as I performed the same experiment he did on day five (which explored the relationship between the number of necrotic T-cells provided to hepatocytes and the number of phagocytosis events). It felt like a culmination of all my training and learning from the past two weeks and I utilised all the skills I had learnt by shadowing and observing Adam performing experiments. In the morning of my ninth day at the University of Birmingham, I observed the cells I had split and cultured the previous Thursday and was pleasantly surprised to find that they were thriving and ready to use for the experiment. Completing this experiment helped me to develop my organisational skills as I carefully followed the protocol Adam had created and balanced different tasks simultaneously.
Overall, my work experience at the University of Birmingham has been a fantastic experience and I am extremely grateful to Dr Zania Stamataki and Adam McGuinness for teaching and supporting me throughout my time here.
* I have provided the list of publications I had the pleasure of reading in case you would also like to immerse yourself into this fascinating field:
Curiosity Killed The Cation 