Researching Virology and its Intricacies from Brain Organoids to Baby Hamster Kidney Cells

Xoshill Saldana, COL ’26, Phoenix, AZ

This summer I had the enriching opportunity of furthering my knowledge and understanding at the Jurado lab in Perlman’s department of Microbiology. As a student who enjoys STEM- I wanted to immerse myself in the ins and outs of working in a lab. As a first year, I decided to take that step and find resources which could allow me to get into a lab. I began working in the Jurado lab in early October and have loved it ever since- which is why I decided to come back in the summer. I gained insight into various techniques, understanding how to operate the equipment, as well as understanding my project. Being in a lab has really allowed me to see the amazing world of science and its intricacies. The beauty of research, its uniqueness and huge impact on humanity and evolution.

The Jurado Lab aims to understand the complex mechanisms of the immune system utilizing emerging viruses to study neonatal health. With these viruses we can overall improve offspring health. What I found poignant and really caught my eye prior to joining this lab was the correlation between the brain and a virus. The harm it does while entering cells inside your body. What I found more fascinating and amazing was the ability of the brain to try to fight the virus off via the presence and expression of antiviral genes (try to fight off the virus so it does not continue to infect other cells).

This summer specifically I had the opportunity to learn new procedures to further understand my project. During the semester I was introduced to techniques such as RNA Extraction, CDNA, and Reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Because I had been doing this for almost the whole year I felt more confident in the summer to do this on my own. Although I had prior knowledge of some techniques I was excited to learn new and longer procedures. My project consisted of researching 3D in vitro brain organoids and looking at the presence of Interferon Regulatory Factor 3(IRF3). IRF3 has many functions, however, in my case IRF3 is a gene that plays a very important role in the immune system’s response during viral infection. These brain organoids are essentially IPS cells that are then clumped together and put into a shaker that is also an incubator to form and mimic a mini brain. My mentor infected these brain organoids with LACV, then homogenized those brains with a chemical called trizol. The remnants were then stored into an eppendorf tube which then I utilized to run my qPCR and a new technique I learned -called plaque assay. I utilized RT-qPCR to understand if there are antiviral genes present within the sample, in this case I was only looking for LACV. Based on the results and data I would proceed with the next steps.

Plaque assay was the other common procedure that quickly became my favorite in the lab. Although it takes a long time I truly enjoy every step. The plaque assay is to see how potent the virus is at certain dilutions. So, we would start by growing cells (I used BHK), once they were grown I would seed them into plates of 6-wells, and then I would infect with supernatant that was collected from cells that were infected with LACV. Next, I would then add agar and plaques would form. Once there were plaques formed, I then would stain with purple violet (purple liquid), and then count plaques. The plaques were an indication that the virus (LACV) had gone into the cells and successfully infected as well as replicated. Depending on the dilution you would either see more or less plaques. I also did a bit of imaging and staining. Which is another technique I learned that was looking at slices of the organoids under a microscope. I stained it with different colors to identify different regions and different cells within the organoid. I truly enjoyed it!

Being in research especially over the summer has shown me that science is not a smooth ride. The process will have a lot of barriers, sometimes even failure-but, it is with these experiences that truly allow a person to learn and find room to refine any weaknesses. I am glad to have had the summer I did and had the ability to gain new insight into the world of research.

This is part of a series of posts by recipients of the 2023 Career Services Summer Funding Grant. We’ve asked funding recipients to reflect on their summer experiences and talk about the industries in which they spent their summer. You can read the entire series here

By Career Services
Career Services