Second Year Summer

Dr Gwendolen Reilly‘s Group

Gwen’s group works on bone tissue engineering, musculoskeletal cell mechanobiology and orthopaedic biomaterials. Previously, the group had shown that Oscillatory Fluid Flow induced mineral deposition in murine pro-osteoblasts. This paper formed the basis of my first funded research project, with the aim of optimising parameters for bone tissue engineering.

10102016-cellculturearea

Fig. 1. The only photo I took in the lab. (Sterile environment in cell culture labs!)

The cell type I used was MLO-A5 pro-osteoblasts/pre-osteocytes in passages 28-33 (relatively late but acceptable as “trials”). Without prior experience, I had a hectic first week of training. In the first week, I was trained to operate under the cell culture hood, fume hood, make up culture medium and stains, passage cells (to include, detaching, putting into a centrifuge, counting, and replacing them into T-25/T-75 or 6-well culture plates for experiments), operate the microplate reader and analyse the data.

As with the experiment itself, we placed the well plates in a portable incubator, which was then placed on a rocker platform. Depending on the experiments, alternative culture media enhanced with Ascorbic Acid or different concentrations of FBS were applied; OFF exposure regime varied upon frequencies of rocking and rocking duration/rest time. Analyses were carried out to quantify viable cells and mineralised cells, by staining with alamarBlue, incubating and extracting for microplate reader analysis, washing, fixing, staining with alizarin Red, washing off the excess, then destaining on a shaker (in circular motion as opposed to the side-by-side rocker) and extracting for another microplate reader analysis. (Phew!) Statistical tests were carried out to look at the significance of results (ie. whether they were random or distinct events).

Outside of the lab, I had the chance to attend weekly research seminars where post-docs and PhD students present their work, often as a rehearsal of their conference presentations. At Kroto Research Institute, researchers were from experimental backgrounds, bringing up interesting discoveries and creations of biomaterials and introducing many different experimental techniques with the equipments at the institute. Contrarily, at the Insigneo Institute, academics were theoreticians who focused on computational modelling of whole organs. This was a larger seminar where experienced academics did scruntinise and question the presenters on the spot. I was fortunate to observe on the side how the senior academics view certain issues and how one should respond to queries while retaining one’s composure. Seminars from both institutes showcased different approaches of medical research using engineering methods, which further strengthened my desire to work in this area.

Back story: Gwen was one of the few Tissue engineers who host a wet lab. Whilst I approached her in my first year for a short research placement – without luck, I have eventually come to this group in a less straightforward way. I was originally awarded the eagerly awaited Wellcome Trust Vacation Scholarship, which is, obviously, a very large funding body and would be a strong support on my CV. On the day when the award letter came through, my original supervisor called me in for a quick chat, and said that he had resigned and could not carry on the project with me. Extremely emotional, he had asked for me a departmental award which matched up with the WT scholarship, and referred me to Gwen. Indeed the experiment I did at Gwen’s was very similar to what I would have done with the original supervisor, with the exception of the cell type used (one being mouse bone cells, and the original one being vascular cells). This has turned out to be an exceptional opportunity though in spite of the preceding surprises, because I had the chance to attend research seminars at both sites, whereas the former supervisor was only a faculty in one of them. With this placement I was exposed to a wider spectrum of research groups (through seminars), but a smaller and more intact team of experimentalists. Nothing as planned, but with more fun.

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