Sartorius eBook - 28
THE FUTURE OF DRUG DISCOVERY: LIVE 3D CELL ANALYSIS
Phase-contrast imaging dynamically monitors iPSC generation and colony emergence over time, and
fluorescent surface labeling confirms pluripotency
A.
Day 7
Day 12
Figure 4: Visualization of iPSC generation from neonatal
Day 17
B.
foreskin fibroblasts using phase-contrast imaging and ICC
(SSEA4-AF-488 and OCT4-AF-594). iPSC formation was
induced from neonatal foreskin fibroblasts by treating with
CytoTune-iPS Sendai Virus Reprogramming Kit (Yamanaka
Factor-containing virus). (A) Whole well images (4X)
demonstrate the emergence of distinct iPSC clones on
Day 12 after addition of reagent. (B) Isolated colonies fixed
and stained with antibodies targeting pluripotency markers
SSEA4- AF-488 and OCT4-AF-594 confirm pluripotency of
iPSCs.
for innovative research, but it also brings fresh
to iPSC differentiation, helps ensure confidence
technical challenges and new requirements for
in results.
cell QC. Despite demonstrated success in the
QC monitoring with live-cell analysis
technique, conversion rates vary from 0.00001-
provides real-time data over extended
1%, making cell reprogramming a relatively
time frames, allowing researchers to detect
rare event that is difficult to detect and confirm.
and characterize extremely rare events.
Being a dynamic process, programming may
Furthermore, the ability to capture the whole
take several weeks or even months to complete,
plate means users can visualize reprogramming
and without visualizing all the cells over the
events, such as the emergence of colonies,
entire time course, events could be missed
and ensure quality before establishing clones.
or misinterpreted, wasting valuable time and
Colony development can be tracked using
resources. Ultimately, the continual monitoring of
label-free imaging, which provides colony
cells, from somatic cell reprogramming through
morphology, size, and growth characteristics
28 |
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Sartorius eBook
Table of Contents for the Digital Edition of Sartorius eBook
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