Author(s): João L. V. Monteiro de Barros

Presentation: poster

The GL261 cell line is a universally used model system to study Glioblastoma Multiforme (GBM), an extremely aggressive type of human brain cancer with a mortality rate near 100%. Brain cancer research on GBM usually relies on a specific GL261 cell phenotype, known as adherent cells (AC). Previous work showed these cells often mutate during normal cell passaging protocols. Here, we examined different passaging rates as a potential selective pressure that alters AC cell phenotype and behavior, thus potentially drifting their genome further from normal GBM cells. We artificially selected for slow- and fast- growing AC cells to test whether they would show similar cellular densities at the end of each passaging time compared to a control group. Our results revealed that slow-growing cells reached 80% confluency during a 5-day passaging rate, as expected, after 3 passages (15 days). However, fast-growing cells that experienced a 1-day passaging rate did not exhibit 80% confluency within a day and rapidly underwent drastic phenotypic changes. This study highlights how important consistent passaging rates for GL261 cell culture is and calls for more specific cell splitting protocols. Otherwise, potentially life-saving brain cancer research is compromised by unintentional selection that changes the experimental cells.

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