There are many perks to working in an undergraduate lab. You get to make great networking connections, work on different projects, and be a part of cutting edge research. I had a great project that worked on bone-marrow derived stem cells (MSCs) that we successfully differentiated into endothelial cells (ECs). This was very exciting for me to be a part of as it was my first year working in a lab, outside of the long hours of lab duty you're assigned to as a biology major. Nevertheless the technique that we used was a nice blend of old practices mixed with some new ideas.
To differentiate our MSCs we used growth factors like FGF, EGF and VEGF to coax them along the path of turning into endothelial cells. They also were plated with different combinations of basal lamina components to assist them in their transition. We had previously differentiated MSCs in vivo with the help of all of the cross-talk between the cells. However, this is what made our task interesting to me. Our goal was to try to determine the key players that were working within the body to differentiate stem cells to whatever cell type was needed, in our case endothelial cells. So after we decided on the components we wanted to test on the cells, we had to decide which model to use: suspension or coating. If you've ever seen the hashtag #overlyhonestmethods, then I think this would fall nicely into that category. We were trying for the suspension model, but since the gel didn't quite set-up we had a “trap” model. This is basically to say we trapped our cells somewhere between the plate and our newly created network. Nevertheless this “new technique” seemed to work out great as our cells differentiated nicely into ECs. This was tested via X-gal staining with a lovely blue to show we had positive cells.
However, our next goal was to implant the cells to see if they could actually form vasculature in the body. It was great that we had positive results, but if it couldn't translate into something therapeutically significant then the excitement would quickly diminish for me. The goal here was simply to see if their dual-nature of being part stem cell and part endothelial cell would allow them to hone in on the site of injury and also start the repair. So we needed an in vivo model to test out our new hypothesis. When looking at our options, we decided to use luciferin. Now, this is where my inner child/geek kind of exploded. As a kid I was fascinated with lightening bugs and their tiny little glowing bodies. I would run around catching them for hours, while trying to figure out how in the world they were able to glow without being plugged in. They apparently seemed to catch the eye of the scientific community as well, as we now use their glow in cell culture. You see, it was great to have positive cells, but we needed to watch their progression inside the body which is not possible with staining. Therefore the new goal was to transfect our differentiated MSCs with luciferase and watch their movement inside the body.
While there are other options to watch living cells, like RFP or GFP, with luciferin we did not need to use a special microscope to view the cells. This was an added bonus for me, since you had to have a key to get in to use the very expensive fluorescent microscope, and I did not. So again this left us with a great model for viewing our living cells, without having to hassle with scheduling for the microscope. The results are still pending, but it was definitely cool to get to work with one of my favorite childhood toys while still being a part of cutting edge research. I would say that makes for a great day any day.
Vanderbilt Class of 2014
Deanna is an intern at Goldbio during the summer of 2013.
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