By Tom Keeley, King’s College London, UK
I’ve spent the last three years looking for a better way to recreate and observe cell responses in the laboratory. This has been the focus for my PhD, ‘Defining an in vitro model of normoxia and its implications for nitric oxide signalling’, which I’ve been researching in the Vascular Biology Lab in the British Heart Foundation Centre of Research Excellence at King’s College London.
Scientists routinely use room-air conditions to study cells at the bench, but we wanted to know if physiological oxygen levels would provide more meaningful results. Our research is showing that signalling pathways can be seen more clearly at physiological oxygen levels, so I’m hoping that scientists will increasingly adopt normoxia as standard rather than as a specialist technique. While room-air studies can definitely be meaningful, it’s much better to replicate the conditions in the body if you’re looking at complex cell situations or developing new drugs.
I usually cycle to work, which can be hairy: I’m from Cornwall, so London can take a bit of getting used to! The first thing I’ll do when I arrive is head for the cell culture suite. I keep a vigorous log of what I’m growing, which usually dictates what I’ll be doing for the rest of the day. If I’m starting a new culture, I’ll need to get the petri dishes to 5% dissolved oxygen before they go in the chamber, which takes a couple of hours, so I’ll use the time to analyse some previous samples. I’ve been working with umbilical cells, because they provide a good model for vascular cells throughout the body. They also offer a balance of both male and female donors, and a good mix of ethnicities.
There’s no such thing as a typical day here. If I’m doing a 12-hour time course, I’ll need to be in at 7am, and I won’t go home until well after 7pm. Some days, I may spend time with my supervisor, talking about how things are going and where to go next with my research. I might also meet up with an undergraduate I’m responsible for; we design lab-based projects for them, which are, of course, intended to benefit our own research! I also travel quite a bit. Not long ago I was visiting a lab in Austria, and I’ll be attending a conference in San Francisco soon.
Part of my work involves feeding back to Baker Ruskinn, the manufacturer of the specialist lab equipment I use for creating normoxic culture conditions. They particularly value end-user feedback, especially as we’re using their equipment differently: we’ve adapted their hypoxic chamber to replicate normoxia instead. We’ve had a lot of support from Baker Ruskinn – they placed equipment with us for a year so I could get the preliminary data together for my PhD funding application. They understand the science, and have been very willing to invest in us without being at all demanding in return.
Since my research is funded by British Heart Foundation, I’ve been focused on cardiovascular therapies. However, the implications of my research are very broad – they could be very relevant for cancer treatments, for example. My findings could have huge implications for the development of new pharmaceuticals, and we might get more meaningful results if pre-clinical drug testing were conducted at the right oxygen level, which could help to get more new drugs onto the market.
My PhD is pretty much complete now, so I’m looking for funding for a further 3 years’ post-doctoral work to continue this line of research. My plan is to be a career academic because I relish the freedom to follow my ideas. For example, I didn’t go into my PhD with any solid objectives or hypotheses; it evolved a lot over the 3 years, enabling me to pursue the areas we wanted, driven by my interests.
That said, research can sometimes be difficult. The hours can be long, and you have to be prepared for the fact that an experiment may not work, your cells may inexplicably die, or your hypothesis may not pan out. You definitely need patience and perseverance. But it’s fantastic when you do figure things out – these personal rewards are exactly why I love science.
My 5 top tips for anybody that’s working towards their PhD are:
- The three P’s are essential; Patience, Persistence and Perseverance. Many of my experiments did not work first time and required significant optimisation before any meaningful data could be obtained. Troubleshooting is fundamental, and requires a great deal of patience and persistence.
- Be open-minded and follow curiosity. The final thesis may be considerably different than your initial hypothesis, which you may have even disproven completely. Keeping an open mind is essential to progress and developing new hypothesises.
- Enjoy the journey. It can be difficult at times but doing a PhD is a real experience and you get to do great things and go to fantastic places.
- If your deciding whether or not to pursue a PhD, be realistic about the challenges and expectations; both your own and your supervisors expectations of you. It is not for everyone and you will find it difficult if your heart is not truly in it.
- Don’t be afraid to be ambitious and creative. Scientific advancement is based on novel, original and sometimes bizarre ideas, so don’t be afraid to discuss these with your supervisor and anyone else who will listen.