High-Content High-Throughput Assays and iPSC Neuronal Cultures

It is no secret that the human brain is in incredibly complex organ that researchers are just beginning to understand. However, just because the brain is shrouded in complexity doesn't mean that methods do not exist that can test certain behaviors in specific situations. This is what Sirenko et al. from Texas A&M have shown with their recent study using high-content high-throughput assays and Human Pluripotent Stem Cells (iPSC) to study neurotoxicity in relation to pharmaceutical drugs. This is a particularly important area of investigation since there have been many neural diseases that have been attributed to chemical exposure, both pre and post natal. To help understand this and to eventually develop a method that could help prevent these types of chemicals from ever reaching humans, much attention has been paid to neurotoxicity and pharmaceutical treatment. However, until now there have been significant obstacles in the way of being able to do this kind of research. For example, the sheer number and immense interconnectivity of brain cells makes it incredibly difficult to replicate in a lab setting. Also, cell maturity and the highly differentiated states found in humans are not easily repeated in cell-based models.

This is where iPSCs enter the stage. They exhibit the function and maturity of human neurons and this makes them an incredibly effective tool for trying to understand neurotoxicity. This, combined with high-content imaging, has helped Sirenko and her colleagues come up with a reliable and reproducible way for measuring neurotoxicity. They measured the effects of chemicals on brain cells by studying mitochondrial membrane potential using the JC-10™ indicator. What helps make this indicator so effective and reliable is that it requires just one step staining. This dramatically reduces the risk of damaging the cells under study to the point where their behavior is affected, which helps maintain the integrity of the results and increases their reliability.

Developing drugs that effectively treat what they aim to treat is already a significant challenge facing the pharmaceutical industry, but ensuring that these drugs have no harmful side effects is another one that must be given equal attention. The method of testing neurotoxicity developed in this study gives researchers the tools they need to be able to accurately understand the effects of drugs on the brain. The availability of high-content imaging was key to confirming the validity of this method. The indicator used, JC-10™, plays a vital role in this because of the robust and vibrant results it produces. Thanks to some improvements over previous indicators, its signal to background ratio is significantly higher, which ensures more robust results. In an environment like the brain where tiny adjustments can makes serious impacts, it is important to be able to rely on the images being presented. JC-10™ helps make this happen and is a big reason why this study was able to confirm its results. Progress made in this investigation will go a long way to helping the medical community in developing safe and effective drugs.

References

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1. Sirenko, Oksana, et al. "High-content high-throughput assays for characterizing the viability and morphology of human iPSC-derived neuronal cultures." Assay and drug development technologies 12.9-10 (2014): 536-547.