Gain Critical Insights from Advanced Cell Models with Real-Time Analysis - 18

Real-Time Analysis of Advanced Cell Models | ADVANCED IN VITRO MODELING

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Figure 4.

Concentration response curves were constructed using an area under curve (AUC)
analysis of the time-course data (Figure 4). Inhibition of multi-spheroid growth
caused by Lapatinib and ZK164015 aligned with the known expression profile of
receptors targeted by these agents. The dual EGRF and HER2 tyrosine kinase inhibitor Lapatinib caused a concentration dependent inhibition of SK-BR-3 and BT-474
multi-spheroid growth, while the estrogen receptor (ER) antagonist ZK164015 was
a potent inhibitor of MCF7 multi-spheroid growth with little or no effect on multispheroids devoid of ER expression. The DNA topoisomerase inhibitor, CMP, caused
comparable inhibition of growth across all multi-spheroids types.
Herceptin-induced ADCC in HER2-Positive Multi-spheroids
Herceptin-induced antibody-dependent cell-mediated cytotoxicity (ADCC) was
evaluated in HER2-positive multi-spheroids. In Figure 5, on the following page,
tumor multi-spheroids either stably expressing nuclear restricted RFP (NR) or
cytoplasmic GFP (CyG) were co-cultured with peripheral blood mononuclear cells
(PBMCs) (E:T, 5:1) and treated with Herceptin, a monoclonal antibody targeting
HER2 receptors. Multi-spheroid proliferation and immune cell-mediated cytotoxicity were quantified kinetically using the IncuCyte S3 metric, fluorescence intensity
within brightfield boundary that does not require masking of fluorescent tumor
multi-spheroid.
Herceptin induced a concentration-dependent cytotoxic effect in HER2-positive
multi-spheroids (SKOV3 and BT-474) but not HER2-negative (MCF7) multi-spheroids.

Figure 4. Perform robust and reproducible pharmacological analysis. A panel of four tumor
breast cell lines were co-cultured with NHDFs on a layer of Matrigel for 3 days to allow
multi-spheroids to form prior to treatment. Concentration response curves represent area
under curve (AUC) analysis of the time-course data (0 -7 or 0 -10 days) post-treatment. Note,
differential compound pharmacology across cell types.

| January, 2019

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

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