Pipetting treatments into cell culture media and mixing

Author: Addison McLaughlin | Major: Biomedical Engineering | Semester: Summer 2023

Background

My name is Addison McLaughlin and I am going into my junior year of biomedical engineering. This summer concluded my first year of research. This year, I studied the PIEZO 1 ion channel. I began this study in the beginning of 2023 and have continued it throughout the past 7 months. The objective of the study was to examine the effects of silencing a mechanosensitive calcium ion channel, PIEZO 1. Included in this was studying the effects of YODA 1 on the osteogenic differentiation of mesenchymal stem cells (MSCs). YODA 1 is a chemical compound that mimics PIEZO 1 in mice and humans. It does this by reducing the mechanical threshold for PIEZO 1 activation, which in turn increases the rate of osteogenic differentiation in MSCs.

Project Execution

Throughout the semester, I have worked to optimize the protocol for YODA 1. This entailed varying the amount of time that the cells were exposed to YODA 1 and the concentration at which YODA 1 was present. I also began my work studying the PIEZO 1 gene. siRNA transfection was performed on MSCs to silence the PIEZO 1 gene and real time polymerase chain reaction (qPCR) was performed to confirm the silencing.

Trouble Shooting

I ran into many problems throughout my research experience. Though the cells were meant to be adhered to the wells, hyper deposition of collagen caused the cells to begin to peel off at the end of the differentiation period, so I had to be especially careful when fixing the cells that I did not aspirate out or dislodge the cells. Another problem I ran into was obtaining different results than what was expected in the second portion of my PIEZO 1 project. I also continued to have issues with the Alizarin Red staining of several experiments that had been treated with YODA 1, where there were few differences between it and the control group, but there were other signs that differentiation had occurred.

Mentorship

My mentor is Dr. Rebekah Margaret Samsonraj. I came to learn about her lab when she was my professor for Introduction to Biomedical Engineering and I joined her lab part way through the fall semester of 2022. She had several projects in the works when I joined her lab. She and I worked together to decide on which one would be best for me, that is how we landed on my chosen topic. My mentor guided me through obstacles and failures in my research by helping me troubleshoot problems as they occurred. She also always encouraged me to read more about my chosen topic and further my understanding of what specifically was happening with the YODA 1 and what each step of the PCR protocol was meant to do.

Conclusion and Acknowledgements

I was assisted by two NWACC students, Will Dockery and Brandon Prado. Will and Brandon helped me with the culturing of the hMSCs and with the staining and analyzing of the results. I am truly grateful for everyone who helped me this past semester. This past semester has been amazing. Through all that I have learned, I was able to vastly improve my problem solving and perseverance skills, as well as my time management skills. I took the problems step by step to try and figure it all out. I plan to continue working in Dr. Samsonraj’s lab throughout the rest of my time at the University of Arkansas, and then I plan to attend graduate school with a focus in tissue engineering.