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 ability to detect hESC colonies was first demonstrated using in vitro fertilization (IVF) plates. Due to the relatively large growth area (2.9 cm2), only the central area where most of the colonies were seeded could be considered (Fig. 3A). By comparing images from day 3 of cultures with those acquired 24 h after seeding, it was possible to assess the growth of the colonies and create striking visual representations of this very dynamic system. The same principle was applied to hESCs growing in the microfabricated bioreactor (Fig. 3B). In contrast to the IVF case, the small dimensions of the culture chamber made it possible to monitor growth based on images of the whole culture area. This enabled determining the response of cells to perfusion: colonies were found to migrate, merge, or even wash out on rare occasions. These results were obtained using intermittent imaging. This approach was next applied to fully automated imaging of mESCs cultured in the reactor for long periods (5 days). The results showed that the image-processing method was able to detect mESC colonies accurately despite the prevalence of artifacts (Fig. 3C). Based on this detection, the confluency of the culture (i.e., the fraction of the culture area occupied by cells) was determined for the duration of the culture. Interestingly, the mean and standard deviation across three trials were relatively low (26%), demonstrating good reproducibility (Fig. 3D).
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.