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Comparative Study
. 2010 Jun 17:10:43.
doi: 10.1186/1472-6750-10-43.

Accurate non-invasive image-based cytotoxicity assays for cultured cells

Patricia Marqués-Gallego  1 , Hans den Dulk, Claude Backendorf, Jaap Brouwer, Jan Reedijk, Julian F Burke
Affiliations
Comparative Study

Accurate non-invasive image-based cytotoxicity assays for cultured cells

Patricia Marqués-Gallego et al. BMC Biotechnol. .

Abstract

Background: The CloneSelect Imager system is an image-based visualisation system for cell growth assessment. Traditionally cell proliferation is measured with the colorimetric MTT assay.

Results: Here we show that both the CloneSelect Imager and the MTT approach result in comparable EC50 values when assaying the cytotoxicity of cisplatin and oxaliplatin on various cell lines. However, the image-based technique was found non-invasive, considerably quicker and more accurate than the MTT assay.

Conclusions: This new image-based technique has the potential to replace the cumbersome MTT assay when fast, unbiased and high-throughput cytotoxicity assays are requested.

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Figures

Figure 1
Figure 1
Cell confluence (%) vs incubation time with cisplatin in the A2780R cells; only results of six wells are plotted for clarity.
Figure 2
Figure 2
Cell confluence (in %) of the A2780R cells treated with cisplatin (rows from A to D), or oxaliplatin (rows from E to H) after 72 h of incubation. First two columns (1 and 2) are non-treated cells. The compounds concentration is increasing from well 3 to well 12 (from the left to the right: 8.47 nM, 25.4 nM, 76.2 nM, 0.229 μM, 0.686 μM, 2.06 μM, 6.17 μM, 18.5 μM, 55.6 μM, and 167 μM).
Figure 3
Figure 3
Bland-Altman plot (pEC50 value difference CloneSelect Imager-MTT vs average) showing the comparison of the pEC50 values produced by cisplatin in: (A) the A2780 cells, (B) the A2780R cells, after 48 h of incubation, tested with CloneSelect Imager system and MTT assay. The limits of agreement (bias ± 1.96 SD) are denoted with horizontal solid lines.
Figure 4
Figure 4
Comparison of the dose response plot of cisplatin after 72 h in the A2780R cells. Both plots display cell viability relative to non-treated cells vs -log [cisplatin]. (A) measure with the MTT assay and (B) measure with the CloneSelect Imager system. The pEC50 value is denoted at the crossing of the solid lines. The concentration of cisplatin used (8.47 nM, 25.4 nM, 76.2 nM, 0.229 μM, 0.686 μM, 2.06 μM, 6.17 μM, 18.5 μM, 55.6 μM, and 167 μM) plotted as -log C vs cell viability. The error bars indicate the accuracy and reproducibility.
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