Nanion Technologies
SOT 2018 Exhibitor Session
San Antonio, March 12th 2018

Innovations for cell monitoringin safety and toxicity assays
Dr. Elena Dragicevic
Senior Scientist / Sales Manager at Nanion Technologies

AgendaCell Monitoring Approaches:• Cell proliferation monitoring• Hepatotoxicity monitoring• Cancer growth monitoring

Cardiomyocyte attachment andtoxicity monitoring via impedance
SyncroPatch 384/768 PE
ion channels in cell lines and iPS CMs
The SyncroPatch 384/768 PE
with Optical Stimulation

CE96

Long-term and short-term monitoring for chronic and acute effects

controled environment

long-term stableconsumables

dual readout fromthe SAME cells

Referenceelectrode

Impedance /EFPelectrodeCE96

Cell proliferation monitored via impedance on CardioExcyte 96
Cell growth

Cell deathCHO cells proliferation monitoring (72h)

Effects of Escin and Triton X on CHO cells (>100h)

CE96

Hepatotoxicity monitoring via impedance on CardioExcyte 96
Effects of increasing concentrations of paracetamol on base impedanceof monocyte-derived hepatocyte-like (MH) cells (>80h monitoring)CE96

Investigating therapy response/resistance ofTherapeutic schedule to analyse the effect of chemotherapy on
mammary carcinoma cell lines
in vitroendogenously induced animals?
Orthotopic Tumor Model

Experimental workflow

Implantation

9 ca.
days6 days

V = 0.5 cm3

V = 0.5 cm3CAF Chemotherapy (i.p.)
Group 1

dissection

Group 2dissection

100 mg/kg Cyclophosphamide
5 mg/kg Doxorubicin (Adriamycin)
100 mg/kg 5-Fluouracil

CAF
Group 3dissection
CAF

Murine H8N8 mammary carcinoma cells exhibit aplastic epithelial or mesenchymal phenotype,validated by in vitro immunofluorescence stainings.

G1 - Untreated: Untreated Control; dissected after tumor growth to 0,5 cm3G2 - Residual: Treated with CAF; dissected at minimal tumor volume
G3 - Relapse: Treated with CAF; dissected after tumor regrowth to 0,5 cm315

Investigating therapy response/resistance of
mammary carcinoma cell lines in vitro
Transparent NSP®-96
sensor plates

Murine
H8N8cells:
immortal
mammary carcinoma cell line withtumor stem cell properties.

H8N8T25 flask

10x

H8N8T3.2CE96

10x10x10xH8N8 T3.2 cells represent arecurrent tumor variant. H8N8 T3.2cells were established from a solid
breast tumor that received a CAF
clinical regimen in vivo.

Poster P374: 'In Vitro Investigations ofAdjuvant Chemotherapy for Preventionof Breast Cancer Tumor Recurrence' on
Tuesday March 13th. Data courtesy of Dr.
Oliver Reinhardt, Dr. Frauke Alves,
Translational Molecular Imaging Group,
Max-Plank Instiute of Experimental
Medicine, Göttingen, Germany

CAF Chemotherapy (i.p.)
100 mg/kg Cyclophosphamide
5 mg/kg Doxorubicin (Adriamycin)
100 mg/kg 5-Fluouracil

CE96Cancer growth monitoring via impedance on CardioExcyte 96

Impedance readout (CardioExcyte 96)

Image-based live-cell analysis readout

H8N8
H8N8T3.2

tumor cell
doubling time (h)60,040,020,00,0Neg. Control
H8N8
H8N8T3.2

CE96

Investigating therapy response/resistance ofmammary carcinoma cell lines in vitro

Transparent NSP-96
sensor plates

H8N8T3.2

H8N8

T25 flask

10x10x10x10x

Impedance readout (CardioExcyte 96)

Image-based live-cell analysis readout

AgendaCell Monitoring Approaches:• Cell proliferation monitoring• Hepatotoxicity monitoring• Cancer growth monitoring

Cardiomyocyte attachment andtoxicity monitoring via impedance
SyncroPatch 384/768 PE
ion channels in cell lines and iPS CMs
The SyncroPatch 384/768 PEwith Optical Stimulation

CE96

CardioExcyte 96 Hybrid Measurements: Mode of Operation

controled environment

Start up

EFPswitch

Impedancelong-term stableconsumables

dual readout fromthe SAME cells

Independent data files for each recording mode.
EFP and IMP recordings of the same cells.

CE96

Cardiomyocyte attachment and toxicity monitored via impedanceon CardioExcyte 96
Development of beat rateand EFP amplitude of iPSC CMs

Chronic effect of Nifedipine (>24 h)

AgendaCell Monitoring Approaches:• Cell proliferation monitoring• Hepatotoxicity monitoring• Cancer growth monitoring

Cardiomyocyte attachment andtoxicity monitoring via impedance
SyncroPatch 384/768 PE
ion channels in cell lines and iPS CMs
The SyncroPatch 384/768 PE
with Optical Stimulation

PE

Automated patch clamp technology
from manual work to high throughput state-of-the art automation
Port-a-Patch50
data points / dayPatchliner
600

data points / day

SyncroPatch 384/768PE
20,000 / 40,000
data points / day

SyncroPatch 384/768PE

6-20

compounds / day

75-300compounds / day

2.5k-10k / 5k-20k
compounds / dayPEThe SyncroPatch 384/768 PE
the 4th generation APC

Giga-seal patch clamping
384 or 768 recording channels
Up to 20,000/40,000 data points
per day

Made in Germany @Unattended operation 4-6 hours

1x 4x 8x
R: low€ 0,16 - 0,65 per data point

R: medium

Ultra flexible: Can be applied in
ALL phases of drug discovery

R: highSyncroPatch 384 PE – consumables NPC-384
Variable hole number and size
Partial run (any 32 wells per run)Partial plates can be run without using the whole chipPE

One protocol takes care of many ion channels
Nav1.5 („INa“)Kv11.1 (hERG – „IKr“)

Kv4.3-KChip2.2 („Ito“)
221

Cav1.2/b2/a2d („Ica, L“)

Kv7.1-KCNE1 („IKs“)

Kir2.1 („IK1“)
2

1

1

2
33

PEIon Channels of the CIPA Initiative – on ONE plate

PEDifferent (iPS CM) cells on one 384-well plate
Vendor iPSC-CMs

The SyncroPatch 384/768 PE
the 4th generation APC

Made in Germany @Ncardia Cor.4U

CHO Nav1.7

PEInter-platform and Inter-site Comparison••CiPA cpds were tested on medium and high throughput APC devices
3 Nanion sites contribute IC50 data on 6 ion channels* Risk of TdP** Chantest

Exemplary results for 3 cpds on hERG and Nav1.5

ChannelhERG

Nav1.5Cpd

IC50
Literature**[µM]

Highesttest
conc.
[µM]

Ambienttemp.

Physiol.
temp.

Ambient
temp.

Ambienttemp.

Physiol.
temp.

IC50[µM]

IC50
[µM]

IC50
[µM]

IC50[µM]

IC50[µM]

Sotalol*

111300160151

192

217

71.4

Diltiazem13.2

100

13.9

15.6

4.1

8.4

5.7

Quinidine*

0.72

101.02

0.92

1.54

1.02

0.78

Sotalol*

1180

300>300>300>300>300>300

Diltiazem

22.4

100

44.4

44.5

25.2

13.3

22.0

Quinidine*

14.6

10>10>10>10>10>10➢ Strong consistency across platforms and sites

PEHTS pharmacology: current-clamp on iPSC Cardiomyocytes

AP raw trace: CTR vs Nifedipine

Online Analysis APD 90

Made in Germany @Cor.4U
Online Analysis APD90 and APD50

AgendaCell Monitoring Approaches:• Cell proliferation monitoring• Hepatotoxicity monitoring• Cancer growth monitoring

Cardiomyocyte attachment andtoxicity monitoring via impedance
SyncroPatch 384/768 PE
ion channels in cell lines and iPS CMsThe SyncroPatch 384/768 PEwith Optical Stimulation

PEThe SyncroPatch 384/768 PE with Optical Stimulation• First Prototype:• 96 blue LEDs (470 nm) below the Chip
• Constant stimulation at selected intensity possible• Stimulation at selected frequency (e.g. 1 Hz) possible

PEThe SyncroPatch 384/768 PE with Optical Stimulation
Example Traces

Depolarization Via ChR2: Nav1.5-ChR2• Proof of Concept:• CHO cells expressing ChR2 and Nav1.5 were used

• Cells were measured in Current Clamp (CC)• - 90 mV were achieved by LFVC (Low Frequency Voltage Clamp)• 1 ms Light Pulse at 2Hz (470 nM) was used for stimulation

Cells: ChR2 + Nav1.5 in CHO provided byThank you!
SyncroPatch 384/768PE.

CellProviders
The CardioExcyte 96.

Nanion PE team

Nanion Booth #534

Nanion CE team