Comparison of Peripheral Blood Mononuclear Cell (PBMC) Isolation Methods: Ficoll-Paque vs SepMate​

Emylee working at lab bench

Students

2021
College

Faculty

Project Summary

During my PURM experience in the Carpenter lab, I was privileged to collaborate with and study under a wonderful team of scientists and clinicians. Under the leadership of Dr. Erica Carpenter, the Carpenter lab focuses on liquid biopsy in cancer research. We are creating a biobank that includes samples across different cancer types and timepoints, as well as developing assays to improve early diagnoses, disease monitoring, and response to therapy.

One of the sample types our lab collects is buffy coat, which contains peripheral blood mononuclear cells (PBMCs). PBMCs are isolated from whole blood and can be used to evaluate an immune response in cancer patients. However, the traditional Ficoll-based isolation process is arduous and time consuming. StemCell Technologies created SepMate tubes, which include an insert filter that simplifies and shortens the buffy coat isolation protocol. The goal of my project was to compare the traditional Ficoll-based isolation method with the SepMate method and to evaluate whether or not our lab could switch to the shorter SepMate protocol. I did so by comparing the cell count (the absolute number of cells harvested) and cell viability (the percentage of harvested cells that are live) with a Z2 cell counter and a flow cytometer, respectively.

Because my major, Health and Societies, is mostly social science based, I had very little lab science experience before doing research in the Carpenter lab. Over the course of the summer, I learned many laboratory techniques, including how to use a centrifuge and a pipetter, how to isolate plasma, serum, and buffy coat from human whole blood, how to establish and maintain cancer cell lines, and flow cytometry. Beyond the bench, I shadowed several physicians and learned about different medical specialties, including medical oncology, radiation oncology, and pulmonology. This type of immersive education is like nothing else I’ve experienced. Further, my colleagues served as mentors and continue to give guidance and support in my education.