Automated and online characterization of adherent cell culture growth in a microfabricated bioreactor.
Bottom Line: While suspension culture processes benefit from decades of development of instrumented bioreactors, adherent cultures are typically performed in static, noninstrumented flasks and well-plates.A machine learning-based algorithm enabled the specific detection of one cell type within a co-culture setting, such as human embryonic stem cells against the background of fibroblast cells.In addition, the algorithm did not confuse image artifacts resulting from microfabrication, such as scratches on surfaces, or dust particles, with cellular features.
Affiliation: Department of Biochemical Engineering, University College London, London, UK Centre for Mathematics and Physics in the Life Sciences and Experimental Biology, University College London, London, UK.Show MeSH
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Mentions: The use of a microfabricated device as both a culture and imaging chamber requires the following key components (Fig. 1A): flow control for culture medium exchange, environmental control to maintain optimal temperature for growth, and an imaging system for monitoring. The implementation of these three components depends on the monitoring strategy. Intermittent monitoring allows the transport of the microfabricated device from its controlled environment (e.g., incubator) to the microscope for image acquisition. In contrast, online monitoring is achieved by having the device permanently positioned on top of a microscope stage and thus requires a suitable setup to maintain optimal growth conditions. Online monitoring offers obvious advantages, such as high image sampling rates, flexible monitoring schedules, reduced contamination risks (no need to transfer to and from an incubator), and a higher degree of automation (and thus reduced user interaction). Its main limitation lies in the number of devices that can be monitored at once with one microscope. To demonstrate the applicability of our imaging and automation routines, we tested both monitoring concepts.
Affiliation: Department of Biochemical Engineering, University College London, London, UK Centre for Mathematics and Physics in the Life Sciences and Experimental Biology, University College London, London, UK.