Page 11 - Live-cellanalysis handbook

Cell Culture Quality Control Assays

Real-time monitoring and analysis of cell culture
conditions and techniques

A large number of variables exist that alter the growth and Each of these controllable variables can adversely affect the
function of cells in culture. Many sources of variability are results and interpretation of data obtained from downstream
largely uncontrollable because they are inherent to the stochastic assays leading to lost time and a waste of valuable, and at times,
processes in biological systems. Other factors can be identified and costly reagents. A key element in controlling adverse variables is
controlled. Some key controllable factors include: to standardize on objective metrics, thereby eliminating human
subjectivity and interpretation.
• Poor CO incubator performance due to lack of calibration and
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stability of temperature, humidity and CO over time. Advantages of performing quality control of cell cultures via
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• Non-quantitative and/or inconsistent procedures for feeding continuous monitoring
and splitting cell cultures prior to running cell-based assays
including inconsistent limits of cell density and feeding The IncuCyte® real-time live-cell analysis system provides a
schedules, inconsistent cell density at the time of assay and label-free, non-invasive method for monitoring cells directly in
changes to cell morphology. the incubator. In sharp contrast to manual monitoring of cell
culture processes, quality control monitoring with real-time live-
• Alterations in media components due to lot-to-lot differences cell analysis automates data capture and cell assessment (Figure
and alteration of concentrations over time due to degradation. 1). Cells can be monitored around-the-clock and at precise,
• Differences in cell culture growth surfaces such as variability regularly scheduled sampling intervals. In addition to supporting
among suppliers, vessel surface treatments and lot-to-lot decisions for current culture processes, historical information can
variability. be retrieved months or years later for comparison of cell lines and
culture growth characteristics.
• Biological issues resulting from growing cells in continuous
culture for extended periods of time which in turn increases the
risk of phenotypic drift, contamination by infectious agents and
cross contamination with other cell types.

Manual Cell Culture Monitoring
Initial cell Assessments made at random Cell utilization decisions
density— locations with variable including timing—
subjective frequencies— qualitative and subjective
operator dependent

Seed vessel Monitor growth Utilize cells TIME

Initial cell Assessments made at precise Split, treatment, harvest or
density— locations with scheduled assay decisions based on
measured for sampling intervals— review of both morphology
every well objective growth curves and quantitative confluence
Live-Cell Analysis and morphology data measurements
support decisions

Figure 1. Comparison of cell culture monitoring methods. Vertical arrows represent interaction points with the cell culture using manual
methods and real-time live-cell analysis. Subjective decisions made during manual monitoring result in variability for cell-based assays.

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