Gene expression profiling
as OCT4, NANOG, NODAL, SOX2, TERT,
DNMT3B, and REX1 were up-regulated
in the iPSCs clones compared to in their
parental fibroblasts, whereas fibroblastspecific genes such as CD44 were
down-regulated in the iPSCs (Figure
7A). Hierarchical cluster analysis clearly
distinguished the iPSCs from their
parental fibroblasts and demonstrated
that the iPSCs derived in this study
shared expression patterns similar
to those of iPSCs derived from BJ
fibroblasts and cord blood CD34+ cells,
as well as H9 ESCs (Figure 7B).
A
B
Fold Change
(iPSCs vs Fibroblasts)
To demonstrate that the isolated iPSC
clones express a broader panel of
pluripotent genes, RNA was isolated
from all six fibroblast lines and their
reprogrammed iPSCs. After cDNA
synthesis, gene expression levels were
measured using the QuantStudio™ 12K
Flex Real-Time PCR System and the
TaqMan® OpenArray® Human Stem
Cell Panel, which contains 609 welldefined genes validated as markers
for pluripotency, plus 22 endogenous
control genes. Gene expression profiling
showed that pluripotency genes such
22
21
20
19
18
17
16
15
14
13
12
11
10
9
8
7
6
5
4
3
2
1
0
-1
-2
-3
-4
-5
-6
-7
OCT4*
NANOG NODAL*
PD-1
Ctrl-2
SOX2*
TERT*
Ctrl-1
LIN28* DNMT3B REX1*
PD-2
MSA
CD44
CULTURE
ENGINEERING
DIFFERENTIATION
CHARACTERIZATION
PD-1 Fibro
Ctrl-1 Fibro
PD-2 Fibro
MSA Fibro
PD-3 Fibro
Ctrl-2 Fibro
BJ Fibro
BJ Fibro-2
PD-1 iPSC
Ctrl-2 iPSC
Ctrl-1 iPSC
PD-2 iPSC
MSA iPSC
PD-3 iPSC
BJ iPSC
Episomal iPSC
H9 iPSC
PD-3
*No expression detected in fibroblasts—Ct value of 36 used for comparison
Figure 7. Gene expression profiles clearly distinguish iPSCs from their parental fibroblasts. iPSCs were cultured in KSR-supplemented iPSC medium on MEFs for 4 days,
then cells were harvested using TrypLE ™ enzyme and stored in RNAlater® solution before RNA isolation using the RNAqueous®-4PCR Kit and cDNA synthesis using SuperScript®
VILO™ Master Mix. Gene expression levels were quantified using the QuantStudio™ 12K Flex Real-Time PCR System and TaqMan® OpenArray® Human Stem Cell Panel. Expression
levels of selected well-described pluripotency markers are shown in A relative to parental fibroblasts. Heat maps of iPS cells and parental fibroblasts (B) show distinct
expression patterns similar to those of other fibroblasts and stem cells. BJ iPSCs: iPSCs derived from BJ fibroblasts using the CytoTune®-iPS Sendai Reprogramming Kit.
Episomal iPSC: Gibco® Episomal hiPSC Line derived from cord blood CD34+ progenitors with seven episomally expressed factors.
20 Life Technologies | Parkinson's cell model
