Performing cell culture for chondrogenic differentiation

Author: Vitali Maldonado | Major: Biomedical Engineering

During the Fall 2021 semester, I worked with my research advisor Dr. Rebekah Margaret Samsonraj from the department of biomedical engineering in testing the effects of epigenetic agents on mesenchymal stem cell (MSC) chondrogenic differentiation.

MSCs are cells that have the capacity to become cartilage, fat, or bone cells, inducing regenerative effects at the site of application. Moreover, osteoarthritis is a common disease affecting 32 million adults in the U.S. (Initiative, 2020) This disease mainly affects cartilage, which has poor healing capacity on its own. Nevertheless, MSCs showed multiple positive outcomes in past clinical trials for osteoarthritis (see clinicaltrials.gov), revealing that these cells are a promising treatment for this disease. However, more effective differentiation techniques for these cells are required. This could be achieved by adding epigenetic agents to their differentiation media.
I chose this research topic with the help of my advisor. We started by evaluating the current need in the regenerative medicine field. We assessed the fact that, in some cases, adding substances to the media in which MSCs are cultured, can lead to cells that are more successful in treating a variety of diseases, including osteoarthritis. However, these substances are still under research and more trials are needed in this area to determine which substance is the best in improving MSC differentiation. Moreover, epigenetic agents can lead to positive results without altering the cell DNA and its main functions. Therefore, my research focused on testing these agents on MSC chondrogenic differentiation. During this experience, I learned general aseptic techniques in laboratory work, how to prepare a solution with different solute concentrations, and how to fix the cells for RNA isolation and histology. Additionally, I got better at pipetting, team communication, and professional writing.

Problems were not absent since I had to face two major ones. The first one was during the experimental setup of one plate. I had to seed a treatment group and a control group. But after adding the cells and centrifuging the plate, the pellets were not visible. Consequently, future chondrogenic media change could not be performed, and the cells would die. This plate was discarded later, and cells on a new plate were successfully seeded. The second problem faced was in the RNA reading. One of the groups did not generate enough data and the differentiation for that group had to be performed again.

Dr. Rebekah Margaret Samsonraj was always there to show me how to perform an experiment the first time I had to do it. She carefully explained the steps and physically demonstrated how to perform them. The graduate students also helped me succeed in the multiple procedures and data collection. They assisted me with cell seeding and RNA isolation and reading.

Nevertheless, this is not the end of this research for me. I plan to continue working on improving MSC differentiation with epigenetic agents in the remaining semesters of my undergraduate degree. Testing combinations of different substances and in different concentrations is how I plan to continue this project. Moreover, in my future career, I want to continue working on improving regenerative medicine treatments and therapies.

References

Initiative, U. S. B. a. J. (2020). The Burden of Musculoskeletal Diseases in the United States (BMUS). In.