Sartorius eBook - 31

THE FUTURE OF DRUG DISCOVERY: LIVE 3D CELL ANALYSIS

need for standardization in culture, and assay

of spheroid formation. The ability to visualize

protocols and analysis.

the growth and development of spheroids, and

Live-cell monitoring offers a solution

measure their size, shape, and uniformity also

to the complicated workflows of 3D cell

allows researchers to evaluate optimal growth

culture and analysis. Using live-cell imaging

conditions, for example, the type of ECM and use

to monitor and QC spheroid development,

of co-cultures (Figure 6).

scientists can improve their understanding
Live-cell monitoring of spheroid development and optimal growth conditions with
brightfield imaging
A.

DF-BF image + BF mask outline
b.  + 2.5% Matrigel

a.  - Matrigel

c.  Largest BF area x 104
8 μm 2
MDA-MB-231
+ MG (2.5%)
6
MDA-MB-231

4

2

B.

0

24

48

72
96
Time (h)

120

144

+ BF mask outline
c.  Total BF area x 105

b.  + NHDF

a. SK-BR-3

9 μm 2 / image

SK-BR-3

6
+ NHDF
3

0

0

48

96
Time (h)

144

192

Figure 6: Visualization of single and multi-spheroid development using brightfield optics.
(A) MDA-MB 231 cells were seeded in 96-well round bottom

(B) SK-BR-3 breast cancer cells were seeded in 96-well flat

ULA plates in the absence or presence of Matrigel® (4K cells/

bottom plates on a bed of Matrigel®, in mono- or co-culture

well, 2.5% Matrigel® v/v) and imaged over 6 days. Incucyte®

with normal human dermal fibroblasts (NHDF) (1:1 ratio, 1K

extended depth of focus (DF-BF) images (3 days post seeding)

cells/well). Incucyte® DF-BF images (8 days post seeding)

are shown with yellow outline mask. All images captured at

are shown with yellow outline mask.  (a) In the absence of

4X magnification. (a) MDA-MB-231 cells fail to form compact

NHDF, SK-BR-3 cells fail to form compact multi-spheroids.

spheroids in the absence of Matrigel®. (b) Compact spheroids

(b) NHDF encourage the formation of more compact

form in the presence of Matrigel®.

aggregates. (c) Time-course show kinetic quantification

(c) Corresponding time-

course plot (Largest BF Area) revealed the increased growth of

and influence of NHDF on the size of aggregates.

spheroids in the presence of Matrigel®.
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Sartorius eBook

Table of Contents for the Digital Edition of Sartorius eBook

Contents
Sartorius eBook - 1
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