BioTek July 2019 - 19

Figure 3. High contrast brightfield cell counts over time relative to confluence.
A comparison of NIH3T3 direct cell counts and percent confluence over time
demonstrate the different characteristics of the two cell growth metrics. Label
free cell counts indicate robust logarithmic cell growth up to full confluence
(dashed line).

Table 1. Gen 5 Image Analysis software setting. Image preprocessing
improves contrast reduces background. Cellular analysis parameters and
advanced analysis parameters are set to optimize identification of each cell
for accurate cell counts.

Comparison of Cell Proliferation Profiles
Across Three Cell Types

Figure 4. Label-free direct cell count profiles for NIH3T3, HeLa and HCT116.
Cells were imaged every 2 hours and culture growth was monitored for 5 days
or until cell counts plateaued.

NIH3T3, HeLa, and HCT116 cells were seeded at low
densities and incubated in a BioSpa 8 for five days.
High contrast brightfield cell counts were used to
define proliferation profiles for each cell type (Figure
4). All cells exhibited robust logarithmic growth under these conditions up to full confluence.

Cercosporamide is a MAPKinteracting kinase (Mnk)
that is a potent and selective inhibitor of the translation initiation factor 4E (elF4E)6. Recent studies have
reported cercosporamide inhibits malignant tumor
outgrowth, including HCT116 colon carcinoma
xenograft tumors6,7. To explore the effect of doxorubicin and cercosporamide on NIH3T3, HeLa and
HCT116 proliferation, cells were treated with eight
drug concentrations in a 96-well fomat and monitored for five days (Figure 5). Kinetic cell proliferation profiles demonstrate the differential response
to doxorubicin and cercosporamide across the three
cell types (Figure 6). NIH3T3 cells were not affected by cercosporamide at any of the concentrations

Measuring Effect of Anti-proliferation
Compounds on Cell Growth Rates
Kinetic cell proliferation studies enable quantitative evaluation of cancer therapeutics. Doxorubicin
is an anthracycline antitumor antibiotic that interrupts cell division by intercalating DNA and inhibiting macromolecular biosynthesis4,5. It is commonly
used in the treatment of a broad range of cancers4.

BioTek July 2019

Table of Contents for the Digital Edition of BioTek July 2019