Cell cultures are not as stable as you think

According to a team of bioscientists from KAUST, the culture environments of different types of cells growing in culture vessels or Petri dishes vary greatly from each other and from the conditions in the living body, which could lead to the “crisis reproducibility” in biomedical research. This finding follows constant monitoring of three different cell types, including human pluripotent stem cells, for three days. This research and others could lead to improved standards and protocols for laboratory cell culture experiments.

The inability to monitor and control cell culture conditions has made it very difficult for researchers to replicate experiments, which is crucial for confirming the significance and accuracy of scientific findings.

“We are currently investigating the functional consequences of environmental instability in cell culture by analyzing gene expression and epigenetic and metabolic changes in cells cultured under well-defined conditions,” says Li.

The team’s future research will aim to develop recommendations on how to improve culture conditions for different cell types. Meanwhile, the researchers suggest that modifications to existing batch culture protocols – by reducing cell density, for example, or adapting culture vessels – could help limit the changes to acceptable ranges. Commercially available systems could also be used to continuously dilute cultures with fresh medium or automatically add gases or adjust medium acidity to maintain cellular environments.

A team of researchers led by marine ecologist Carlos Duarte and bioscientist Mo Li monitored the environment of cells grown in flasks placed in a controlled incubator – a standard cell culture method called batch culture – over a period of three days. Three different cell types were used: human pluripotent stem cells, a cancer cell line and a type of white blood cell. Optical sensors were attached to some of the flasks belonging to each cell type to monitor changes in dissolved oxygen and carbon dioxide levels as the cells grew. Additional flasks were removed every eight hours and then discarded after measuring cell growth rates and culture acidity.

“We were surprised to find that the batch-cultured cellular environments were several different magnitudes from their native environments in the human body,” said Ph.D. student Samhan Alsolami.

The extent of changes in cultures varied by cell type, but generally as cell density increased, the amount of dissolved oxygen in the cell medium decreased while carbon dioxide increased. This in turn increased the acidity of the surrounding medium. The extent to which these changes occurred in each cell type is likely due to differences in cell growth rates and metabolism. These changes will affect cellular processes.

“Scientists are still uncovering the factors that contribute to maintaining an ideal cell culture environment to accurately mimic the human body,” said researcher Shannon Klein. “But it’s not an easy task and requires parallel advances in technology and engineering,” she says.

Republished with kind permission from KAUST. Photo: (Left to right) Shannon G. Klein and Mo Li, as well as Samhan M. Alsolami and Carlos Duarte (not shown) aim to develop recommendations on how to improve culture conditions for different cell types. Credit: © 2022 KAUST

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