Applications
Cell-Based Applications >> Cytotoxicity
Cell Adhesion
Cytotoxicity Assay Features
  • Robust, label-free measurement of cardiomyocyte contractility and toxicity
  • Continuous, real-time monitoring of acute and long-term cytotoxicity
  • Universal assay compatible with cell lines, primary cells and stem cells
  • Information-rich kinetic data predicts mode of toxicity
  • High throughput, label-free assay: 96-, 384- and 1536-well formats
  • Run assay and collect data within incubator at 37°C and 95% humidity
Compound Cytotoxicity Screening
Figure 1 - Cytotoxic compounds give distinct BIND Profiles, depending on their mechanism of action. HeLa cells were plated in triplicate on 384-well BIND CA2 biosensors at 20k cells/well and incubated overnight. The next day, cells were treated with 100µM concentrations of compounds known to act via different mechanisms of cytotoxicity.

Figure 2 - Hela cells were plated on BIND optical biosensors and treated with 33µM concentrations of three different steroidal estrogen receptor modulators (SERMs). Biosensors were placed on a BIND Reader located within a 37°C/95% humidity incubator and PWVs monitored once a minute for 20 hours. The inset highlights the first 60 minutes of compound induced toxicity. Reference for Tamoxifen, 4-hydroxy tamoxifen and Raloxifene toxicity: Favoni and de Cupis, Trends Pharmacol Sci. 19:406 (1998).
BIND® technology provides a platform for highly sensitive readouts of cell death. As cells undergo necrosis or apoptosis the morphological changes incurred can be measured in real-time as concentration-dependent changes in Peak Wavelength Value (PWV). Toxins that work through distinct mechanisms of action (e.g. DNA damaging agents, protein synthesis inhibitors, microtubule inhibitors) generate distinct kinetic profiles on BIND® Readers (Figure 1) that serve as signatures for that particular cell death mechanism. The robustness of the BIND Reader allows its placement within a 37°C/95% humidity incubator for continuous monitoring of cytotoxicity.


BIND cytotoxicity assays can differentiate not only the cytotoxic potential between different compound classes, but also can detect differences in toxicity between compounds within the same class. Tamoxifen, a known cytotoxic compound, produces significant cellular BIND responses seen as a sharp negative PWV shift with a continued negative plateau (Figure 2). 4-hydroxy tamoxifen, a metabolite of tamoxifen, is known to cause increased toxicity. It produces larger and more rapid changes in BIND PWVs (Figure 2 inset). Raloxifene, also a steroidal estrogen receptor modulator, has dramatically reduced side effects and cytotoxicity when compared to tamoxifen. In keeping with its lower clinical toxicity, raloxifene induces very minor PWV changes supporting the use of BIND cytotoxicity assays as an early indicator of potential compound toxicity.
Cardiomyocyte Toxicity Assay
Figure 3 - Axiogenesis Cor.At® mESC-derived cardiomyocytes (Axiogenesis/Lonza) were plated at 20k/well in a 384-well BIND optical biosensors and cultured for 72 hours. Potassium Chloride (KCl) was added to a final concentration of 50mM to half of the wells. PWVs measured 4 times/second.

Figure 4 - Cor.At mESC-derived cardiomyocytes (Axiogenesis/Lonza) were plated at 5k/well in a 384-well BIND biosensor and cultured for 24 hours. PWVs were measured 4 times/second on a BIND reader for 2 minutes. Various concentrations of doxorubicin were added and cells incubated at 37°C for 17 hours. PWVs were then measured 4 times/second for 5 minutes.
The advent of embryonic stem cell (ESC)-derived cardiomyocytes enables in vitro toxicity assays on biorelevant cell types that could serve as reliable predictors of in vivo toxicity. The BIND cardiomyocyte toxicity application provides for multiple high value readouts in a single high-throughput, label-free assay.

ESC-derived cardiomyocytes can be grown directly on BIND biosensors and display a healthy, beating phenotype after several days in culture. When placed on the BIND reader, the beating phenotype of cardiomyocytes is detected as oscillations of positive-to-negative PWV shifts. Rapid, real-time readings can be acquired (up to 4/second) that accurately reflect beating frequency and amplitude (Figure 3).

Doxorubicin, a compound with known cardiotoxic effects, causes dose-dependent disruption of both the frequency and amplitude of beating (Figure 4). While acute effects on beating can be detected using rapid PWV measurements, the ability to continuously monitor beating over hours and even days provides valuable information on longer-term toxicity (Figure 5A). In addition, concentration response curves at any time point can be derived allowing for the accurate measurement of IC50 values (Figure 5B).

The physiological relevance of ESCs in conjunction with the label-free nature and high throughput format of BIND cardiomyocyte toxicity assays, make them a powerful in vitro tool for identifying drugs with cardiotoxic qualities in early stage drug development.

A. Doxorubicin Continuous Toxicity Monitoring

B. Doxorubicin Dose Response Curve

Figure 5 - Cor.At mESC-derived cardiomyocytes (Axiogenesis/Lonza) were plated at 5k/well in a 384-well BIND biosensor and cultured for 24 hours. Cells were treated with various concentrations of doxorubicin or buffer. Biosensors were placed on a BIND reader located within a 37°C incubator and PWVs monitored continuously for 17 hours. A concentration response curve was generated from results taken at 17 hours post doxurubivin addition and the IC50 calculated.