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

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