18

• • • • 

ES

 

 

 

 –

αα

α α

ES cells

50μm

ES

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

Myogenesis

DNA microarray

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

1 EGFP

50µm300µm

EGFP/M-cad/DAPI

M-cad EGFP

EGFP/MHC/DAPI

EGFP MHC

%Area

%Number

EGFP MHC

DAPI

%Number

%Area

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

IαIHABP Merge

50µm

24

IαI Hyaluronan-binding protein (HABP)

Serum-derived hyaluronan-associated protein (SHAP-HA)

TSG6 TNF -stimulated gene 6 product)

SHAP-HA

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

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

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

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

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

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

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

4. ESMACS

Islet1 TuJ1Islet1Merge

100µm

―

Nestin

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

100nM 500nM 1µM

(K252a)

PCR