It’s White Paper Wednesday! Read our featured white paper: Increasing Efficiency in Cell Line Development using the Celigo Imaging Cytometer
In this study, using the CHO cell line, the Celígo Imaging Cytometer was used to monitor transfection efficiency, identify passaging times, optimize media, identify single cells and track clone outgrowth, following previous studies detailing Celígo’s performance in tracking cell lineage and confluence [3, 4]. After transfection, cells were allowed to stabilize, then imaged in bright field and fluorescence to monitor and evaluate transfection efficiencies. As wells grew asynchronously, bright field imaging was used to track which wells needed passaging. As genetically modified isolated cells have decreased growth rates, design of experiment (DoE) methodologies and automated imaging were used for media formulation improvements to increase their growth and survival. Media supplements that have been shown to increase growth were tested with an eight parameter DoE [3 levels, 3 factors, and single output response]. While the identification of single cells in bright field is possible, using the fluorescent marker such as Cell Tracker Green or GFP to monitor the single cells proved advantageous. Wells growing a single cell were subsequently monitored for the formation of a colony, which also allowed for measurement of optimal passaging times of asynchronous clones. Overall, the Celigo Imaging Cytometer demonstrated the utility of automation in the development and monitoring of new CHO-based cell lines for increasing efficiency in cell line development.