Sartorius eBook - 39

THE FUTURE OF DRUG DISCOVERY: LIVE 3D CELL ANALYSIS

tumor breast cell lines; MCF7, MDA-MB-231,

time-courses illustrate a concentration

SK-BR-3 and BT- 474. Cells were co-cultured with

dependent inhibition of ZK164015 and CMP on

NHDFs (1:1 ratio, 1,000 cells/ well of each) and

spheroid size (Figure 4).

multi-spheroids allowed to form for 3 d prior

Visualization of brightfield images support

to treatment (up to 10 d) with standard-of-care

specific cytotoxic effects of Lapatinib on

and cytotoxic agents, Lapatinib, ZK164015, and

SK-BR-3 size and a non-specific cytotoxic effect

Camptothecin (CMP).

of CMP on both MCF7 and SK-BR-3 spheroids

Incucyte®'s real-time, automated vessel

(Figure 5).

views enabled rapid visualization of treatment

Concentration response curves were

effects on multi-spheroid size (Total BF Area).

constructed using an area under curve (AUC)

96-well vessel views of MCF7 multi-spheroids

analysis of the time-course data (Figure 6).

with overlay analysis mask and treatment

Inhibition of multi-spheroid growth caused
Figure 4: Incucyte® S3 vessel views enable rapid
visualization and quantification of treatment
effects. MCF7 cells co-cultured with NHDFs
were seeded in pre-coated (Matrigel) flat
bottom 96-well plates (1:1 ratio, 1,000 cells/
well of each) and multi-spheroids allowed to
form (3 d). Spheroids were then treated with
serial dilutions of known standard of care and
cytotoxic compounds (5 d). Incucyte® microplate

Lapitinib (10
μM)

Treatment

ZK164015
(10 μM)

CMP
(10 μM)

vessel views show effects of treatments on

Controls

multi-spheroid size. Top image shows Brightfield
Object Area (μm2) segmentation mask (yellow)

MCF7-NR + NHDF

5 d post treatment. Bottom image shows the

1

2

3

4

5

6

7

8

9

10

11

individual well Total Brightfield Object Area (μm2)

12

5e5

(y-axis) over time (0-5 d post treatment) (x-axis).

A0

5e5

B
0

5e5

C
0

5e5

D
0

5e5

E
0

5e5

F
0

5e5

G
0

5e5

H
0

5d

Treatment

5d

Lapitinib

5d

5d

5d

ZK164015

5d

5d

5d

CMP

5d

5d

5d

5d

Controls

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| 39


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Sartorius eBook

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