Throughout summer, I had the honor of working in the Neurology department in the Children’s Hospital of Philadelphia as a research assistant under the mentorship of Judith Grinspan and Kelly Jordan-Sciutto. Starting off my summer in the lab, I was taught the basics of what to do in a laboratory setting and the essential techniques of working with culture samples. I learned the importance of sterility when working with in vitro rat models and live cells as well as the various biology and chemistry protocol techniques I would be conducting throughout my time in the lab such as immunocytochemistry, which stains different cells in the cultures with fluorescent protein markers, and immunoblotting, which shows the amounts of certain proteins in the culture samples.
Dr. Grinspan’s lab has recently been interested in antiretroviral drugs and their role on oligodendrocyte maturation. About fifty percent of Human Immunodeficiency Virus (HIV) patients experience a cognitive disorder called HIV-Associated Neurocognitive Disorder (HAND) despite being on antiretroviral drug therapies (ART). These patients have been shown to have a decrease in white matter, which is generated by oligodendrocytes. This information prompted the lab to test the effects of different antiretroviral drugs on oligodendrocytes with in vitro rat models. The ART drug I used, Abacavir, is commonly used on young children in combination with other ART drugs, due to the fact that different ART drugs target different aspects of the HIV replication process. Since it has been known that oligodendrocyte precursor cells (OPCs) can turn into either mature oligodendrocyte cells or astrocyte cells, the goals of my project were to identify if Abacavir was inhibiting the maturation of OPCs into mature oligodendrocyte cells, and if so, whether or not the astrocyte cell line was being activated. To test these questions, I implemented standard biology protocols, immunoblotting and immunocytochemistry, to look at the number of oligodendrocytes and astrocytes in the cultures.
While conducting my research, the lab ran into a few problems that we needed to try and fix in order to move on and obtain significant results. Our immunoblots were not showing up as clear as previously intended, so we had to experiment with the amount of protein added to each well to try and find a balance where we could see the bands on the western blots while not having bands that were too blown out. This process of trial and error within a specific protocol really showed me firsthand how every step of the way with research has to be tested and perfected to be able to generate a list of procedures that will allow you to obtain results that are readable and quantifiable.
The opportunity I had through PURM was one that I probably would not have gotten anywhere else. I was able to be deeply involved with a graduate student’s research project and see the everyday struggles and triumphs that researchers experience. While I had participated in labs in biology class, the experience of being in a professionally run lab was like no other.