Sartorius eBook - 43

THE FUTURE OF DRUG DISCOVERY: LIVE 3D CELL ANALYSIS

1.28e-5 μg ml-1
6.4e-5 μg ml-1
3.2e-4 μg ml-1
1.6e-3 μg ml-1
0.01e μg ml-1
0.04 μg ml-1
0.2 μg ml-1
1 μg ml-1
MS Alone
MS + PBMCs
MS + IgG (1 mg/mL)
MS + AntiCD3/IL2 (10 ng/mL)

SKOV3-NR

BT474-CyG

Red intensity x 106
(RCU x μm2/image)
8

Green intensity x 106
(RCU x μm2/image)
24

6

18

4

12

2

6

Figure 8: Kinetic quantification of ADCC immune
cell killing of HER-2 positive multi-spheroids.
Tumor cells were seeded in flat bottom 96-well

0

0

48

96
144
Time (h)

192

240

0

0

48

96
144
Time (h)

192

240

plates (1,000 cells/well on a bed of Matrigel) and
allowed to form multi-spheroids (MS) for 3 d.
Once formed, MS were co-cultured with freshly
isolated PBMCs (E:T, 5:1) and treated with serial
dilutions of Herceptin. Time-courses show multispheroid death quantified as a loss of fluorescence
intensity within the spheroid brightfield object.

MCF7-NR
Red intensity x 106
(RCU x μm2/image)
24

% of control (FLU intensity AUC)
100
MCF7

18

75

12

50

Concentration response curves to Herceptin show
sensitivity differences between HER2-positive

SKOV3

multi-spheroids (SKOV3 and BT-474). Treatments
targeting T cell populations (Anti-CD3 and IL-2, 10

25

6

BT-474

ng/mL) induced maximal MS cytotoxicity across
all cell types. Data were collected over 10 d at 6 h
intervals. Each data point represents mean ± SEM,

0

0

24 48

72 96 120 144 168
Time (h)

0
-12

-10

-8
Time (h)

-6

-4

n=4 wells.

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Sartorius eBook

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