Recap: Real Time Cell Analysis Workshop.

Thanks for joining us at the Real Time Cell Analysis Workshop at the 2^nd Australian Cancer Metabolism Meeting.

During the workshop, we learnt how the xCelligence from ACEA Bioscience employs non-invasive impedance microelectrodes in automated high-throughput plate formats to maximize the physiological relevance of data extracted from in vitro cellular assays. These instruments enable label-free, real-time monitoring of cell proliferation, cell size/morphology, cell-substrate attachment quality, and cell invasion/migration. By eliminating the time- and labor-intensive steps of traditional methods, RTCA vastly improves efficiency and overall productivity. By continuously providing real-time data in short (minutes to hours) and long (days to weeks) time course experiments, RTCA provides a phenotypic view of cellular behavior at an unprecedented level of detail.

For our experiment, Meso-1 cells (a mesothelioma cell line) were seeded at 3000, 5000, and 10000 cells/well and allowed to grow in regular growth media for 23 hours. 50uM of ICG-001, an inhibitor of TCF/β-catenin-mediated transcription, was then added. Data collection was paused for 4 hours and then collected for an additional 28 hours.

Each well produces an individual data trace which you can see below.

Legend:

From the data we see the following:

1. Addition of ICG-001 results in cell death independent of cell number
2. The biphasic effect of the compound would be lost if the experiment was stopped too early – we would have assumed that the compound does not induce cell much cell death
3. Short term effects, such as morphological and adhesion changes in the first four hours of seeding can be captured
4. Each cell number has a unique Cell Index (CI) proliferation curve, so the curve can be used to identify if the correct cell is being used and also the health of the cell.

These readings are label-free, so the experiment could have been stopped at any time to perform orthogonal assays. The mechanism of action of the compound can also be predicted by the shape of the CI curve and these data can also be used to determine the best time point to run other complementary assays, such as flow cytometry based apoptosis assays.