Being a Masters student isn’t quite what you might expect. Sure there are the standard deadlines, the coursework, the stress, but let’s take a moment to appreciate the actual science going on here.
As you can no doubt see, this particular piece has been written rather close to the deadline for this blog (Sorry Markus!), but I can assure you there is good reason! I wanted to have completed a big as a chunk of my MSci as possible before putting fingers to the keyboard, so I am better able to voice my thoughts. My aim here is to describe my project and explain my thoughts, and I promise to try to keep the complaining to a minimum!.
The goal of my project is to determine how a cell changes during differentiation – the process of a cell turning into a different cell type (Figure 1) – with a specific focus on messenger RNA, or mRNA. To do this, we took a mouse embryonic stem cell line (ESCs), and grew them in culture. A point to note is that these cells want to change. Their whole purpose is to differentiate into all the different cells types one might find within a mouse. This makes it very difficult to culture them, as we want them only to differentiate when we tell them to. Biologists, however, have an answer to this in the form of a compound known as Leukaemia Inhibitory Factor, or LIF. This wonder of biology has the marvellous effect of inhibiting differentiation, thus cells growing in LIF will remain pluripotent.
Figure 1: Stem cells are able to differentiate into multiple types of cells, from muscle to liver cells, though exactly what changes within the cells themselves is somewhat poorly understood. Source
And this is what we did. Two weeks of watching these ESCs be cultured in LIF, and finally we had enough to do the experiment. It’s common practice to made freeze downs of cell cultures as well, so each time the cells were fed whilst being grown, we checked the total number of competent, or living, cells. When we saw there were a lot – I couldn’t see the exact point at which “a lot” happened, but the experienced eyes of my supervisor could – the cells were split, with some continuing to be grown and the rest being frozen. These freeze downs essentially act as a restore point, the same way one might back up computer files, just in case they are needed.
So, having grown the cells we set up the experiment we wished to do, and waited the needed time for the cells to do their thing. Fast forward 72 hours, and we find our first attempt didn’t succeed. The vast majority of cells had died, however they had differentiated like we wanted, clearly we had simply added too many cells to the plate this time. Attempt 2 went much more smoothly, allowing us to harvest the cells for later analysis.
Now, the keen readers among you might have noticed the use of the word “we” a lot so far. This is somewhat of a misnomer. It is “we” in the sense that I, the author, was in the same room. Alas due to the nature of this particular project, I was unable to do any of the physical lab work, and it was carried out by others. This was quite saddening in fact, as all my friends were doing proper lab work, and I was resigned to taking notes whilst watching over their shoulder. Not doing the actual work however, did offer some enlightenment. It taught me how research is not all about physically doing the experiment, but rather it’s about the data collected.
This said, I do feel that research is in part about doing experiments, and I have found it rather challenging to enjoy my project as much as I expected because of it. It has always been my vision that working in a lab is about wearing that not-so-white coat and watching stuff grow, but this project showed me my vision isn’t always the case. Aside from the lack of practical skills, the project has been rather interesting, it introduced me to an area of developmental biology that I didn’t know existed. Challenging though it was, and indeed still is, to understand the content needed for my project, the guidance I have received in the form of my supervisors has helped significantly.
Other than cell cultures, I have also seen how qPCR – a way of quantifying how much of something there is in a sample, such as DNA – works up close, and been part of the data analysis as part of my project. In fact, my project is almost solely about reinterpreting already published data, and to great success. We actually saw something new and exciting! Though you’ll have to wait until the data is published to find out exactly what that is. The contributions that this knowledge could make however is outstanding. For a result so simple, it could change human medicine in the long term, or even how we think of cellular differentiation in the short.
Even though my project was not everything I expected, and the unfortunate fact I am not able to fully participate in the lab work, I do still find research something I enjoy immensely. The topic is interesting, the content continually being updated and improved upon, with my only regret being I personally being unable to participate more directly.