Limitations imposed by non-physiological cell culture practices: characterization and solutions

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Each year thousands of original research papers report results of studies performed on cultured mammalian cells to better understand fundamental biology processes at the level of individual molecules. Almost all of these studies use commercial media that look nothing like human plasma (in which cells are bathed inside the body) and fail to regulate oxygen at the levels found in the body. The failure to mimic normal body conditions when working with human cells affects the reliability of the data we glean from experiments. Data from cell culture is critical not only to understanding normal biology, but also mechanisms of disease. If cells are grown in media with little resemblance to blood and behave differently than in the body, their use as experimental models is compromised. Just this year (2019) there has been attention given to developing cell culture media with compositions very similar to blood plasma, and we have begun to use these. Unfortunately, we have already gathered enough data to know that virtually all of our cell culture results are different in this new media compared to traditional cell culture media upon which the foundation of cell biology has been built. This Discovery Grant proposal focus on identifying and characterizing these differences in the behaviour of cells growing in new media that resemble human blood vs the old commercial media that bear little resemblance to blood. Ultimately, this will contribute to our ability to do better experiments that are more likely to accurately model reproduce in vivo biology and thus to allow better drugs to be developed. We propose investigating gene expression, metabolism, free radical production, and growth under the old conditions and in the new media. Once we have characterized problems with the old cell culture approaches, we will work to develop solutions that help us maintain the right conditions when working with cells in culture. For example, we will investigate the use of a synthetic hemoglobin to aid in oxygen delivery to cells. We will also investigate how to prevent changes in media composition over the course of long experiments. We will continue to develop our existing tools that allow us to visualize mitochondrial form and function inside of cells. Mitochondria play important roles in energy and metabolism, so their health and function are important. This work will be done mainly by MSc and PhD students who will work with me in my research laboratory to gather data and develop new tools. They will attend international conferences to share their discoveries and inventions with other scientists. Undergraduate BSc students and grade 12 students will also work with us to learn laboratory and research skills. Through these opportunities, all these students will progress toward highly skilled professions like medicine and allied health, technical positions in hospitals, universities, and government institutions, industrial or academic research, and teaching.