Viable cell density and culture viability are critical process parameters in cell culture.  Accurate and reproducible quantification of the cells in your process is critical to batch success.  With the evolution of technology, several instruments have emerged on the market that utilize various techniques for cell quantification.  In this study, three instruments were evaluated to quantify cells that presented varying morphologies.  A CHO cell line was used as the standard single-cell suspension culture.  It was compared to a non-uniform, clumpy culture.  Finally, cells grown on microcarriers were also evaluated. The CEDEX currently stands as the gold standard for quantification of single cell suspension culture; however, instrument design and software setup present challenges in the quantification of clumpy cell lines and microcarrier culture.  Using the popular Trypan Blue Exclusion method, the CEDEX captures microscopic images of the cell sample and a pre-designed algorithm accepts items of a certain size as cells and items above and below that size as non-cellular, excluding those in the cell count.  As a result, Clumpy cells are often disregarded by the software as non-cells, and are not quantified accurately.  Microcarriers, on the other hand, can clog the intricate tubing system of the instrument unless they are clarified from the sample first. To address these challenges, the NucleoCounter � 100 and NucleoCounter � 3000 were tested.  The mechanisms of action of these instruments provide a biological advantage that is favorable for the quantification of clumpy and microcarrier cell culture.   The NucleoCounter NC-100 has an integrated fluorescent microscope that detects the dye Propidium Iodide (PI), which binds to DNA.  The details of these instruments, as well as advantages and disadvantages for each cell line are discussed.  Using the data generated from this study, we can make a recommendation for cell quantification for each of these cell types.

Evaluation of alternative instruments for cell quantification of single cell culture, clumpy culture, and microcarrier culture

243rd National Meeting (2012)

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