Gain Critical Insights from Advanced Cell Models with Real-Time Analysis - 13

Real-Time Analysis of Advanced Cell Models | QUALITY CONTROL ASSAYS

and consistency of kinetic assays, it is important
to minimize variations in cell seeding densities
due to pipetting errors during plate seeding
which result in differential growth rates. Measuring confluence before utilizing assay plates
eliminates assay variation, resulting in reproducible conditions.
* Documenting time-dependent variables: Assessment of cell behavior and growth in association with cell treatment times can define experimental conditions to ensure consistent results.

Cell Culture QC Assays: Measuring proliferation to optimize growth conditions and seeding
densities
The IncuCyte system can be used to track cell proliferation over time using label-free phase segmentation (confluence) metrics (Figure 2), allowing for
the assessment of cell growth as well as monitoring
cell culture and assay optimization.

Figure 3. Human umbilical vein endothelial cells (HUVECs) cultured in basal cell media containing growth factors
and supplemented with decreasing amounts of FBS. The optimal concentration of FBS for normal propagation was
determined to be >2.5%.

Figure 3 shows results of a study to assess optimal
levels of fetal bovine serum (FBS) and cell density
using the IncuCyte system.
Live-cell analysis can be used to accurately mask
different morphologies and quantify label-free
proliferation over time, as shown in Figure 4. In
4A and 4B, the variation in cell seeding densitydependent proliferation in two different cell lines,
A172 human glioblastoma and LNCap human
prostate carcinoma, is exemplified. Understanding
cell seeding variation and its effects on proliferation is important for controlling assay variability. As
shown in 4C, images reveal accurate segmentation
of cell types irrespective of morphology.
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| January, 2019

Figure 4. Quantification of A172 human glioblastoma (A) and LNCap human prostate carcinoma (B) growth curves
using label-free confluence analysis. (C) HD-phase contrast images of A549 human lung carcinoma, HT-1080 human
fibrosarcoma and LNCap cells shown without and with confluence mask (orange). Images reveal accurate confluence
masking of all three cell types of varied morphology.

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Gain Critical Insights from Advanced Cell Models with Real-Time Analysis

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