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18
• • • •
ES
–
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αα
α α
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ES cells
50μm
ES
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Telomerase activity ( amol / μL) 0.1 0.2
ES Cells
CD105+ MSCs
ES Cells (Heat block)
CD105+ MSCs (Heat block)
Blood cells (Heat block)
ADSCs (Heat block)
N.D.
N.D.
N.D.
N.D.
0
ADSCs
Blood cells
DAPI CD105TelomeraseCD105+ cells
Merge
10μm
MSCs Telomerase
Differentiation 2011 132 6 669-672p Nana Ninagawa Mesenchymal stem cells originating from ES cells show high telomerase activity and therapeutic benefits
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Myogenesis
DNA microarray
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MyoD
Myf5
M cadherin
Myogenin
GAPDH
ES cells Skeletal muscle cells from MSCs
RT-PCR
50µm
50µm 20µm
C2C12
MSCs
M-cad MHC
8 days6 days
28 days8 days
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1 EGFP
50µm300µm
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EGFP/M-cad/DAPI
M-cad EGFP
EGFP/MHC/DAPI
EGFP MHC
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%Area
%Number
EGFP MHC
DAPI
%Number
%Area
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Percentage of load response phase
Sham Transplantation
%
Inju
red
leg
norm
al l
eg
BioReseach Open Access 2013 8 14 2 4 295-306p Nana Takenaka-Ninagawaa Transplantation of Mesenchymal Stem Cells Derived from ES Cells Promotes Muscle Regeneration and Accelerates Functional Recovery of Injured Skeletal Muscle
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IαIHABP Merge
50µm
24
IαI Hyaluronan-binding protein (HABP)
Serum-derived hyaluronan-associated protein (SHAP-HA)
TSG6 TNF -stimulated gene 6 product)
SHAP-HA
![Page 14: 名大の授業 (NU OCW)#)18$ R S ^yw> ~ •vvv s 7{k xvvF'NTi 7 j •vvvH L@ , e x s*+{ |1Dv p ¢ ¢ ¡ •vvvNT ey b I zx xvvb s w MyoD Myf5 M cadherin Myogenin GAPDH ES cells Skeletal](https://reader035.vdocuments.net/reader035/viewer/2022071006/5fc3813d531b302c321e9676/html5/thumbnails/14.jpg)
cont +Lysate
50µm
** P<0.01
* P<0.05
TSG
6 m
RN
A
Fold
cha
nge
of c
ompa
rativ
e ra
tio
0
2
4
6
8
10
3h 6h 9h 12h 24h 48h
*
Lysate +Control
Incubation hours
******
**Real-time PCR
TSG6 DAPI
LysateMSCs
Lysate
Ann
![Page 15: 名大の授業 (NU OCW)#)18$ R S ^yw> ~ •vvv s 7{k xvvF'NTi 7 j •vvvH L@ , e x s*+{ |1Dv p ¢ ¢ ¡ •vvvNT ey b I zx xvvb s w MyoD Myf5 M cadherin Myogenin GAPDH ES cells Skeletal](https://reader035.vdocuments.net/reader035/viewer/2022071006/5fc3813d531b302c321e9676/html5/thumbnails/15.jpg)
TSG6 KD+lysateCont. +lysate
EGFP/DAPI
50µm
0
10
20
30
40
50
60
70
80
Con. 8 TSG6 KD
Condition of MSCs
Are
a of
set
tled
MSC
s (m
m2 )
P<**
**
TSG6 knockdown (sh mRNA)
MSCs 1. TSG6 2. Myotubes lysate 3. Lysate MSCs
IαI HA SHAP-HA
MSCs TSG6 SHAP-HA 50µm
TSG6
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iPS
iPS
Rat iPSCs
Rat MSCs
100µm
EGFP iPSC
50µm
SSEA1 EGFP DAPI ( )
Nanog DAPI Nanog ( )
50µm
Transfection of lentivirus
(Oct4, Klf4, Sox2, EGFP)
ALP
Leptin r
Lean
Obese
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Nervous tissues
Tuj1 DAPI Tuj1 DAPI EGFP Tuj1 DAPI EGFP
10µm 10µm
Mesenchymal tissues
Endoderm tissues
H-E Collagen II DAPI EGFP DAPI MHC
10µm50µm50µm
H-E Cdx2DAPI EGFP Cdx2DAPI
50µm 50µm
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iPSB
0
0.2
0.4
0.6
0.8
1
1.2
1.4
Adi
pone
ctin
/ G
APD
H m
RN
A
0
0.2
0.4
0.6
0.8
1
1.2
DS/lean DS/obese
PPA
Rγ
/ GA
PDH
mR
NA
0
0.2
0.4
0.6
0.8
1
1.2
1.4
DS/lean DS/obese
Adi
pone
ctin
/ G
APD
H m
RN
A
0
0.2
0.4
0.6
0.8
1
1.2
1.4
DS/lean DS/obese
PPA
Rγ
/ GA
PDH
mR
NA
iPSCs adipose tissue
Lean
Obese
Real-time PCR
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3. ES
Stem Cell Discovery 2012 2 2 70-77p Hitomi Hosoe Investigation of VEGF and PDGF signals in vascular formation by 3D culture models using mouse ES cells
100µm
AteloCell (Collagen gel)
CD105 DAPIFlk1
10μm
Merge
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VEGF+ VEGF+ VEGF+PDGF B PDGF B PDGF B
** * **
VEGF PDGF-B
Stem Cell Discovery 2012 2 2 70-77p Hitomi Hosoe Investigation of VEGF and PDGF signals in vascular formation by 3D culture models using mouse ES cells
VEGF PDGF-B
PECAM1 EphrinB2 Flt4
VEGF
VEGF BDGF-B
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4. ESMACS
Islet1 TuJ1Islet1Merge
100µm
―
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Nestin
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0 0.5 1 1.5 2 2.5 3 3.5 4 4.5
Islet1 mRNA
- HB9 mRNA
no - EF 4d - DCEF 2d - DCEF 4d - Pulse 2d - Pulse
* *
* *
Real-time PCR
Islet1Tuj1 cont Tuj1 Islet1
50µm
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100nM 500nM 1µM
(K252a)
PCR
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