Application Note

Cells: human cardiomyocytes
Tools:
CardioExcyte 96

Assessing cardiotoxic risk of anti-cancer
agents on Nanion’s CardioExcyte 96
University of Göttingen and Nanion Technologies GmbH, Munich.
Human stem cell-derived cardiomyocytes kindly provided by Ncardia.
Results

New anticancer agents have led to higher life expectancy

The impedance signal recorded on the CardioExcyte

for patients surviving cancer. However, treatment-

96 changes as a result of alterations in confluency, cell

related morbidity factors such as cardiac toxicity

contact (morphological shape) and conductivity of

have become important issues for long-term cancer

adherent cells and thereby provides a measure ofsurvivors1-3. Cardiotoxic side effects such as arrhythmia,toxicity. In addition, effects on contractility can also

thromboembolism and myocardial ischemia are common

be measured by monitoring the mean beat (MB). Basewith anti-cancer drugs such as the anthracyclines . This

impedance of hSC-CMs grown on the NSP-96 plates forhas led to the need for a sub-speciality of medicine,

the CardioExcyte 96 was monitored over 170 hours after

cardio-oncology

treatment with paclitaxel (Figure 1A & B).
BAVehicle
5 nM Paclitaxel
50 nM Paclitaxel

patient-derived hiPSC-CMs offers new possibilities for4themeasurementof

electrical

impedance

coupled with human stem cell-derived cardiomyocytes(hSC-CMs) we could confirm the cardiotoxic effects ofpaclitaxel, also known as Taxol, a microtuble stabilizing
drug approved for the treatment of breast, ovarianand lung cancer5. In addition, we investigated different
combinations of cylophosphamide (CP), doxorubicin(DOX) and 5-Fluorouracil (5F) and found that any
combination which included DOX was highly cardiotoxic.

Download more Application Notes from www.nanion.deAmplitude (Ω)

5UsingµMeen

TwnM-80nM

0 24 48 72 96 120 144168
Time (hr)[Paclitaxel]risk of developing cardiovascular complications based
genetic factors4.

-60

C

personalized medicine, being able to assess a patient’s
on their own cells, thus taking into account their own-40

5cell derived cardiomyocytes (hiPSC-CMs) and, indeed,

600
400
2000-200chemotherapy. In addition, advances in human stem

800

2050

to facilitate effective cardiovascular health during

1000

Control
24 hr
48 hr
168 hr4050

risk of new and existing cancer therapies in order

500 nM Paclitaxel
5 µM Paclitaxel
TweeneIt is important to be able to assess the cardiotoxic

Base impedance (Ω)

to fight cancer.

nM

promoteclto

5oncocardiology,hiorcardiovascular health whilst providing the best therapy

Ve

1-3

Base Impedance (% Change)

SummaryVehicle
5 nM Paclitaxel
50 nM Paclitaxel
500 nM Paclitaxel
5 µM PaclitaxelTween

3210
0.0

0.1

0.2

0.3

0.4

0.5

0.6

Time (s)Figure 1: A Increasing concentrations of paclitaxel induce a decreasein base impedance of Cor.4U cells which can be monitoredcontinuously.
Black arrow indicates addition of compound, greyarrows indicate wash with fresh media without compound. Tween(2%) induced 100% cell death and was used as a positive control.
B Paclitaxel causes a concentration- and time-dependent decrease inbase impedance and decrease in amplitude of the mean beat (C).

Application Note
Paclitaxel has been approved for the treatment of ovarian, breast and lung cancer since the 1990’s5. However,
it is also known that paclitaxel is associated with adverse
side effects including cardiotoxicity6. Paclitaxel showeda concentration- and time-dependent effect on base
impedance of hSC-CMs (Figure 1A & B), consistent with achange in cell health. The amplitude of the mean beat
was also reduced indicating an effect on contractility(Figure 1C).

200-401+00.0

0.1

0.2

0.3

0.4

0.5

minimal effects on base impedance were observedPCODOXDOTw Xeen5F

5F+X

5F+X

D+2observed in all combinations containing DOX, whereas

mean beat indicating an effect on contractility.

P

3effects of DOX as a decrease in base impedance was& B). DOX also caused a decrease in amplitude of theC4known to cause cardiotoxic side effects, which can be-80[Drug combination]
VehicleCP
5F
CP + 5F
CP + DOX + 5F
DOX + 5F
DOX
CP + DOX
Tween

5ever, anthracycline antibiotics such as doxorubicin are-60

O

0 24 48 72 96 120 144 168
Time (hr)monly used to treat recurring breast cancer7. How-when using CP or 5F, alone or in combination (Figure 2A-20CAmplitude (Ω)

Control
24 hr
48 hr
168 hr

40

D

1000
800
600
400200DOX
CP + DOX
Tween

The combination therapy of CP, DOX and 5F is com-potentially fatal8. We could confirm the cardiotoxic

B
Base Impedance (% Change)
vehicle
C CP
P+5F

CP + 5F
CP + DOX + 5F
DOX + 5FVehicle
CP5F

Base impedance (Ω)

ATween (2%) induced 100% cell death and was used as a positive control.
B In all combinations containing DOX, a time-dependent decrease inbase impedance and decrease in amplitude of the mean beat (C) wasobserved, whereas only minimal effects were seen where DOX was notpresent.

In summary, the CardioExcyte 96 can be used toinvestigate the cardiotoxic risk of new and existing
cancer therapies. Indeed, with the use of patient-specific
hiPSC-CMs, this offers the potential to personalize the risk
factors of cardiotoxicity of cancer therapeutics basedon genetic and sex variabilities of individual patients. The
impedance signal of the CardioExcyte 96 is a reliable,
non-invasive and accurate measurement for testing

0.6

cardiotoxicity of cancer therapeutics.

Time (s)Figure 2: A Various combinations of therapeutic agents cyclophosphamide (CP; 0.64 µg/ml), doxorubicin (DOX; 0.032 µg/ml) and 5-Fluorouracil(5F; 0.64 µg/ml) caused different effects on base impedance. In all combinations where DOX was present, a cardiotoxic effect was observed.

Impedance measurements

References
1. Bovelli, D., et al. 2010. Annals of Oncology 21 (Suppl. 5): v277–v282
2. Cubbon, R.M., & Lyon, A.R. 2016. IHJ l68: s77 – s85
3. Lenneman, C.G., & Sawyer, D.B. 2016. Circ. Res. 118: 1008-1020
4. Sheng, C.C., et al. 2016. J. Am. Coll. Cardiol. Basic Trans. Science. 1:386–98

Impedance measurements were conducted according
to Nanion’s standard procedures for the CardioExcyte
96. Cor.4U cells were pre-plated on NSP-96 plates andthe base impedance signal allowed to stabilize before
compound added. Compounds were applied in theexternal media. About 2 hours before drug application

5. Weaver, B.A. 2014. MBoC. 25: 2677-2681
6. Marupudi, N.I., et al. 2007. Expert Opin. Drug Saf. 6(5): 609-621 7.
7. Chatterjee, K., et al., 2010. Cardiology. 115:155–162
8. Fukuda, M, et al., 1999. Oncology. 7 (Suppl. 3): 77-81

Methods

the medium was completely removed from the wells and200 μl fresh medium was added. All signals werenormalized to a group of control measurements (n=5-11)
on the same plate. Cardiomyocytes were incubated in
the compound for 48 hours after which the media wasreplaced

Cells

We thank Ncardia for providing the hSC-CMs (Cor.4U)

withfreshmedia

withoutcompound.

Impedance signal was monitored for a total of 190 hours.

and Oliver Reinhardt and Dr. Frauke Alves from theUniversity of Göttingen for performing the experiments.
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