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Candidate cell sources for regenerative medicine include:1. ES cells, 2. Fetal cells, and 3. Adult stem cells.
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Umezawa, A., et al.. Mol. Cell. Biol. 11: 920-927, 1991
Marrow stroma serves a microenvironment or nichefor hematopoietic cells
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Fat Neuron
Bone
Cartilage
Cardiomyocyte
Skeletal myocyte
Default Default With exposure to inducersWith exposure to inducers
Differentiated phenotypes of marrow stromal cells
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ねずみ5匹ねずみ5匹エックス線写真エックス線写真
4 weeks
2 weeks
In vivoIn vivo osteogenic activity in marrow stromal cells osteogenic activity in marrow stromal cells
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PLGA sheets
CollagenCollagenmicrospongemicrosponge
overlay the interstices of overlay the interstices of the fabricated web-likethe fabricated web-like
Generation of bone tissue of desired sizes and shapesSheet-style biodegradable polymer, PLGA (200 um thick) as a scaffold
Hybridization of the PLGAwith collagen microsponge
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The cell-seeded sheets were rolled up around a silicone rod,and cultivated them in vivo for 4 weeks at the subcutaneous tissue.
Cylinder-shaped bone
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Bone knotsbone of more complex shapes
We can control bone shape completely
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Generation of phalanges by the PLGA hybrid sheet with marrow-derived mesenchymal cells.
phalange-like bone
Thumb of the graduate student
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Cell X
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Transplantation of Cell X pelleted micromasses in the nude mice
2W 4W 8W
toluidine blue
HE
x40
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Transplantation of Cell X pelleted micromasses in the nude mice
2W 4W 8W
toluidine blue
HE
x200
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Brain from BoneBrain from Bone
KUSA-A1 cells started to form axon-like long processesby the demethylating agent.
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The transdifferentiation was enhanced by Noggin, an inhibitor of BMPs.
MAP2
Isolated mature osteoblasts have strong in vivo osteogenic activity, and could efficiently convert into functional neurons.
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The stroma-derived neurons started to respond to depolarizing stimuliand neurotransmitter, glutamate, like functional mature neurons.
Differentiation (Special issue on stem cells) 68: 235-244, 2001
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Mesenchymal Cell Lines Summary of Lineage Analysis
KUSA/A KUSA/O CMG 9-15c(MSCs) KUM4 KUM5 KUM5A11E1 KUM9 NRG OP9
Endothelial Cells Flk-1 - - - - - - - - -CD31 - - - - - - - - - -CD144 - + - + - - - - -
Hematopoietic Cells CD34 - ++ - ++ ++ - - ++ ++ -c-kit - - - + - - - - - -Sca-1 +++ ++ +++ +++ +++ + + +++ +++ ++
Mesenchymal Cells CD140 ++ ++ + + ++ ++ ++ ++ +
CD14 - - + - - - - - -CD29 + ++ ++ ++ + + + + + -CD41 - - N.D. + - - - - -CD44 +++ +++ +++ +++ ++ ++ ++ +++ +++ -CD45 - - - - - - + - -CD49b - - + - - - + - -CD49d - - - - - - + - -CD54 - - + - - - + - -CD90 - - - - - - + - -CD102 - - - - - - + - -CD105 - - - - ++ ++ + - - -CD106 - ++ + - + ++ ++ ++ - +Ly-6c ++ + ++ ++ ++ - ++ + ++ +Ly-6g - - - - - - + - - -
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Human Marrow Stromal Cellsreach senescence after a limited number of cell division
H4-1H4-1H4-1H4-1
慶應義塾大学医学部慶應義塾大学医学部血液内科血液内科木崎 昌弘木崎 昌弘福地 由美 福地 由美
H4-1, senescenceH4-1, senescence
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Prolongation of the life span of human marrow stromal cellsby transferring Bmi-1, E6, E7, and TERT
The cells with the extended life span retained the multipotency into osteocytes, chondrocytes, adipocytes, and myocytes in vitro.
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Scheme of the experimetal protocol
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1 week 3 weeks
Recording of action potentialsfrom spontaneously beating cells
Alexa568Human marrow stromal cells with the extended life span
can differentiate into cardiomyocytes
Disorganized
Regular and stabilized
Recording microelectrode
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最近の U7 の誘導率の増加 movie
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EPC-100, EPC-214
Gene transfer
Primary cultureEPCs
Limiting dilution
hTERTE6 (HPV16)E7 (HPV16)
EPC-100 EPC-214
18S(200bp)
E6(397bp)
E7(178bp)
TERT(144bp)
EPC-100 EPC214
Hum
an H
eart
hEPC
100(
RT-)
hEPC
214(
RT-)
hEPC
100
hEPC
214
DW
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GFP labeled
Human cell
5-azacytidine/Oxytocin/-
Adeno-virusFetal mouse
Primary culture of cardiomyocytes
Co-culture
Start beating
2-3 days later
プロトコール
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GFP/h-CardiacTroponin-I/HoechstE
PC-1
00
Hoechst EGFP h-cardiac troponin-I Merge
EPC
-214
Hoechst EGFP Merge
h-cardiac troponin-I
Mouse
EPC-100
EPC-214
h-cardiac troponin-I
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Hoechst/actinin/Connexin 43E
PC-1
00
Hoechst Cx43αactinine Merge
EPC
-214
Hoechst Cx43αactinine Merge
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EPC から記録した活動電位
Pipette
Pipette
EPC
-100
EPC
-214
1sec
50 m
V
2 sec
50 m
V
APD90 MDP AMP Rate Pacemaker(+)
msec mV mV s n
EPC-100 326±12 -55±1 56±7 0.9±0.1 3/17
EPC-214 185 ±16 -48±3 56±3 2.4±0.2 16/22
APD; action potential duration, MDP; Maximum diastolic potential, AMP; amplitude
2sec
50 m
V
1 sec
50 m
V
Pacemaker potential(+) Pacemaker potential(-)
Pacemaker potential(+) Pacemaker potential(-)
Mean ± SE
EPC-100 は同期が強く、活動電位も regular で静止膜電位も深い EPC-214 は同期は弱く、活動電位は培養経過中 (3W) は、歩調取り電位を有した洞結節型細胞
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EPC-100 (2W) Patch clamp Fast Na current was observed in 1/10 cellEPC-214 (2W) Patch clamp Fast Na current was observed in 1/5 cell
EPC
pipette
Patch clamp from post-FACS EPCs
GFP 陽性細胞を FACS にて抽出、その細胞から patch clamp を行った
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Non fusionEP
C-1
00A
EPC
-214
Hoechest EGFP h-cardiac troponin-I Merge
Hoechest EGFP h-cardiac troponin-I Merge
B C D E
F G H I
Feeder cardiomyocyte
Collagen membraneculture dishEPC
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Feeder (+) (-) (+) (-) (+) (-) (+) (-) (+) (-) (+) (-)
Non 5Az Oxytocin Non 5-Az Oxytocin
EPC-100 EPC-214
Indu
ctio
n ra
tio
100
50
40
30
20
10
0
(%)
32%
20%19%
3.5% 4.2% 4.6%
44%
27%
15%
21% 19%
31%
Venus/Hoechist での induction ratio
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CD antigen EPC-100 EPC-214
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Hum
an H
eart
hEPC
100
hEPC
100(
RT-)
hEPC
214
hEPC
214(
RT-)
hEPC
100
hEPC
100(
RT-)
hEPC
214
hEPC
214(
RT-)
Mou
se F
eede
rDW
co-culture (-) (+)Expression of cardiomyocyte-specific gene
Csx/Nkx 2.5 (233bp)
GATA4 (475bp)
hANP (406bp)
hBNP (206bp)
TnT (152bp)
TnI (233bp)
Cardiac Actin (400bp)
MYL2v(382bp)
MLC2α (376bp)
HCN2 (230bp)
18S (200bp)
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心筋誘導までのシェーマ
現在進行中
そろそろ ??
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GFP labeled
Human cell
5-azacytidine/Oxytocin/-
Adeno-virusFetal mouse
Primary culture of cardiomyocytes
Co-culture
Start beating
2-3 days later
共培養のプロトコール
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Csx/Nkx 2.5H
uman
hea
r t
hE-M
SC 1
00
hE-M
SC 1
00 R
T-
hE-M
SC 2
14
hE-M
SC 2
14 R
T-
UB
T-5
UB
T-5
RT-
UB
ET-7
UB
ET-7
RT-
UEE
T-1 2
UEE
T-12
RT-
UET
-13
UET
-13
RT-
DW
Hum
an h
e ar t
hE-M
SC 1
00
hE-M
SC 1
00 R
T-
hE-M
SC 2
14
hE-M
SC 2
14 R
T-
UC
B
UC
B R
T-
PL-9
0
PL-9
0 R
T-
PL-1
12
PL-1
12 R
T-
DW
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400
300
200
100
0
7006005004003002001000290285280275270265260
10008006004002000 30(min.)
20100
心 筋 短 縮 率
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間葉系幹細胞
拡大培養に要求される3つの特性
・安全・安全・迅速・迅速・大量・大量
再生治療
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0 0.5 1 2 10FCS(%)
0
20
40
60
80
100
120
PPL
PLVMSCGM(10%FCS)
UEET-12 無血清培養
改良培地は無血清下でも血清含有培地と同等の増殖性を有する。
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1053210.50FCS(%)
0
50
100
150
200
C
No CoatFibronectin
UEET-12 細胞接着性
接着性の低下は 1 %血清の添加で補われる。
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0 5 10 15 20 25培養日数
1
10
100
1000
10000
100000
#2/X#2/10%DMEM#4/X#4/10%DMEM#5/X#5/10%DMEM
ヒト骨髄由来間葉系幹細胞の初代培養
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p16 INK 4 a /p 14ARF 遺伝子領域
p16 INK 4 a
Cdk4/6サイクリン D
RbE2F
p14 ARF
MDM2(HDM2)
p 53
p 21Cdk2
サイクリン E
細胞増殖
リン酸化 Rb
E2F
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MSCGM X
p16INK4a
β‐tubulin
培地 X ではp16INK4 a 発現上昇がみとめられる
蘇生後の継代 1 1 2 2 3 1 1 2 2 3
0 5 10 15 20 25day
8
10
12
14
16
18
20
22
PDL MCSGM
X
MSC
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なぜ培地 X で p16 の発現が高いのか?
Q
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p16INK4a
β‐tubulin
X + FCS -IT+FCS
-IT -PDGF- b FGF
-EOP -Sel -P,b+FCS-P,b MSC
p16 誘導の主要要因は PDGF,b FGF である。
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p16INK4a
β‐tubulin
X-P,b X-P,b/10%FCS MSCGMPDGF+bFGF(ng/ml) 0 1 10 0 1 10 0 1 10
bFGF,PDGFが p16 発現を誘導する。
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PDGF+b FGF によりp16発現が上昇
なぜ培地 X で p16 の発現が高いのか?Q
A
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MSC bFGF P-AAP-BB
P-AB
+bFGF
P-AA,BBP-AA
P-BBP-ABPDGF
-AAPDGF
-ABPDGF
-BB
A 鎖 B 鎖
β レセプターCD140b
α レセプターCD140 a
Greb2Nck
Ras -GAP
Crk SrcSHP2
PLC-r
PDGF-AA→Competent FactorPDGF-BB→Competent and Proguression Factor
IgG CD140a CD140b
p16
βtubulin
― +MSC の p16 発現は PDGF-B鎖による。
( 細胞 /H10‐8)
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MSC
aFGF
bFGF
EGFVEGFPDGF
PDGF+aFGF
PDGF+bFGF
PDGF+EGFPDGF+VEGF
aFGF+bFGF
aFGF+EGF
aFGF+VEGF
bFGF+EGF
bFGF+VEGFEGF+VEGFHeL
a
p16p16
βtubulinβtubulin
H4-3Normalized Flags Raw Common Description
1.0818335P 2172.2egfr epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)1.1522822P 657.3egfr epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)0.6146532A 337.2egfr epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)0.8937527A 17.8egfr epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)1.0583736A 167.9egfr epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)0.8768445A 358.1egfr ErbB1-S; Human epidermal growth factor receptor precursor (EGFR) mRNA, complete cds.1.0504522A 10.4fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)0.7824624A 21.5fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)0.417403A 9.8fgfr3 fibroblast growth factor receptor 3 (achondroplasia, thanatophoric dwarfism)
0.54518497A 27.2fgfr3 fibroblast growth factor receptor 3 (achondroplasia, thanatophoric dwarfism)1A 38.3fgfr4 fibroblast growth factor receptor 4
1.2821407A 125.7fgfr1 fibroblast growth factor receptor 1 (fms-related tyrosine kinase 2, Pfeiffer syndrome)1P 652.2fgfr1 fibroblast growth factor receptor 1 (fms-related tyrosine kinase 2, Pfeiffer syndrome)
1.4052641A 14.3fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)0.75333506A 59.2fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)1.3886433P 124.5fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)1.1120172A 13.6fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)1.0663294P 2619.3fgfr1 fibroblast growth factor receptor 1 (fms-related tyrosine kinase 2, Pfeiffer syndrome)
1A 11.1fgfr4 fibroblast growth factor receptor 41.1922568A 46.9fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)3.1074321A 73.5fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)
1A 67.3fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)1A 8.2fgfr2 fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)
1.3065747P 3125.7fgfr1 fibroblast growth factor receptor 1 (fms-related tyrosine kinase 2, Pfeiffer syndrome)3.3271298P 4049.1pdgfrb platelet-derived growth factor receptor, beta polypeptide1.0704594P 4990.3pdgfra platelet-derived growth factor receptor, alpha polypeptide1.0662563P 366.9pdgfrl platelet-derived growth factor receptor-like0.8164289A 6.2pdgfra platelet-derived growth factor receptor, alpha polypeptide2.3044996A 403.1pdgfra platelet-derived growth factor receptor, alpha polypeptide0.6091959A 19.5flt4 fms-related tyrosine kinase 41.7572612A 61.6kdr kinase insert domain receptor (a type III receptor tyrosine kinase)
aFGF-R ○bFGF-R ○EGF-R ◎VEGF-R ×PDGF-R ◎
受容体の有無
p16 発現は増殖因子の種類による。
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bPAPBhP
bPAbPBbhP
bPABaEV
PaPEPVabbEbVaEaVEV
-0 20 40 60 80 100 120
mAbs=450nm
MSC äeëùêBàˆéqÇ…ÇÊÇÈëùêBå¯â
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細胞増殖に都合の良い増殖因子ほど p16 の発現を誘導する?
相関係数:
0.700 50 100 150 200 250cell growth
0
20
40
60
80
100
120p16/cell growth
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時間
分裂回数
MSC:MSCGM?毛乳頭細胞: 10%FCS-DMEM表皮角化細胞: 3T3 培養フィーダー乳腺上皮細胞: 3T3 培養フィーダー
MSC:培地 X?毛乳頭細胞:低血清培地表皮角化細胞:無血清培地乳腺上皮細胞:無血清培地
血清培地
増殖因子
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時間
分裂回数
第一世代(血清培地)
第二世代(無血清培地)
第三世代(いやし培地)第三世代(いやし培地)
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増殖因子 ストレス
Ras
Raf
MEK1/2
ERK1/2
Ets1/2
MLKs、 TAKASK1
MKK3/6
p38MAPK
A
B
p16 INK4a
p16ink4a
β-tubulin
X10%FCS
X A B
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間葉系幹細胞
拡大培養に要求される3つの特性
・安全・安全・迅速・迅速・大量・大量
再生治療
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移植細胞の体内動態追跡システム細胞治療において移植された細胞がどこに、どのように存在しているかを知ることは治療評価において重要。現在までに移植細胞の体外検出システムは存在しない。
ドナー細胞およびレシピエントに影響を及ぼさない標識マーカーで細胞をラベリングし、体外画像機器にて検出するシステムを確立する。
【 目的 】
ナノバイオトレースシステム
マーカー;造影剤画像機器; MRI
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Super Paramagnetic Iron Oxide (SPIO)
Feridex 150~250nm Resovist 57nm
T2 短縮効果強い
超常磁性体酸化鉄
1mL 中にフェルカルボトラン 540mg (鉄 24.9mg )添加物: D- マンニトール、乳酸、水酸化ナトリウム
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SPIO 効果 ; T2 短縮(信号低下)
T2 強調 SPIO 造影 T2 強調
•正常肝細胞は鉄コロイドを貪食するので信号が低下する•貪食能のない病変は信号が低下しないので白く浮き出て見える
Labeling
Fe
Fe
FeFe
Fe Fe
Fe FeFe
FeFe Fe
Fe
FeFe
Fe
FeFe
Fe FeFe
FeFe
Fe
Fe
Phagocytosis orPinocytosis
SPIO labeled cell
SPIO
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国内で使用可能な SPIO リゾビストを用い、
ヒト骨髄間質細胞への導入、および生体内に
おける長期間の観察を行えるか?
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10% FBS DMEM0.1% Risovist
Berlin Blue stain
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Pinocytotic vesicle1500x
TEM
Risovist particles
SPIO incubation for 2 days
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Ex vivo Relaxometry
MR spectrometerMinispec NMS-120 (Bruker Japan)
0
10
20
30
40
50
60
70
80
90
100
controlRisovist
Control
Risovist
ControlRisovist
cells/mL
1.44x106 2.88x106 5.75x106 1.15x107 2.3x107
T2 weight T2 value
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In vivo MRI analysis
5x106 cells labeled with Risovist
Suspend into 50µL medium
Injection into the leg muscle
MRI analysis at various time points
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3 hours 2 weeks 4 weeks
MRIT2W
H-E
BB
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Risovist
Polymer sheet
Implantation with scaffoldsT2W
2 weeks after transplantation
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Risovist labeled cell onto PLGA sheet
H-E BB
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H-E B-B
Eight weeks after injectionT2W
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細胞を用いたムコ多糖症 VII 型マウスに対する中枢神経病変に対する細胞治療・遺伝子治療
(大橋十也、島田隆)
出生前診断・胎児治療(末岡浩、林聡)
先天性代謝異常症に対する造血幹細胞移植
日本国における現状(加藤俊一)
細胞治療におけるヒト細胞の安全性の検討ならびにバリデーション
(梅澤明弘)
リソソームの電子顕微鏡写真
リソソーム病とは
遺伝的要因によるリソソーム酵素活性の低下
リソソーム内に分解されない分子が発生
これがリソソーム外に排泄されず進行的に蓄積する。
細胞機能が低下し、臨床症状を呈する。
de Duve (1966)
ムコ多糖の分解酵素の欠損グリコサミノグリカン分子(デルマタン硫酸・ヘパラン硫酸・ケラタン硫酸・コンドロイチン硫酸・ヒアルロン酸)がリソソーム内に蓄積することにより、細胞・組織・器官の機能異常が生じる。
特異顔貌、骨形成異常、聴覚・視覚・気道・心血管・関節可動域の異常、精神発達遅滞
先天性代謝異常症に対する幹細胞治療法の開発
ホルモン(インスリン)産生細胞を用いた細胞治療(中村公俊、宮本薫)
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Transduction of human GUSB gene into BMS cells with a retroviral vector
MND Human GUSB LTRSV40 Neo
GUSB activity
5.9×10
(nmol/hr/mg)
1.9×106
GUSB staining
KUSA/A1 (before)
KUSA/HBG (after)
äeéÌç◊ñEÇÃGUSBäàê´
840.3
141.8 70.6 24.8 61.6126.1
0.0
0100200300400500600700800900
1000
ê_åoä≤ç◊ñE
DEC Th1KUSA/A1
H162 H123 3521
U/mg protein î|ó{è„ê¥ÇÃGUSB äàê´
34.2
1.3 2.5
13.7
1.0 3.90.0
05
1015202530354045
ê_åoä≤ç◊ñE
DEC Th1UET13
KUSA/A1‰`ë—ååcontrol
U/mg protein神経幹細胞と骨髄間葉系幹細胞のGUSB 活性値の比較
福原康之、奥山虎之らによる
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Histochemical detection of GUSB-positive cells in treated mouse brain
Bar equals 200 mm
Cortex
Subventricle
Olfactory bulb
Distribution of GUSB activity in the brain 2 or 8 wk after
transplantation
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Reduction of GAGs contents after transplantation
T-CS
0
100
200
300
400
500
600
UntreatedMPS VIITreatedMPS VIINormalUntreatedMPS VIITreatedMPS VIINormal
UntreatedMPS VIITreatedMPS VIINormal
4wk 8wk 16wk
nmol/hr/mg
HA
0102030405060708090
100
UntreatedMPS VIITreatedMPS VIINormalUntreatedMPS VIITreatedMPS VIINormal
UntreatedMPS VIITreatedMPS VIINormal
4wk 8wk 16wk
nmol/hr/mg
*
* * **** **
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Improvement of cognitive function
Acquisition (d0-d5)
Probe test (d6)
NormalTreated MPS VII
Untreated MPS VII
Target quadrant
Mean non-target quadrants
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先天性代謝異常疾患に対する造血幹細胞移植の実施状況梅沢班・辻 班・日本小児血液学会合同調査梅沢明弘、辻 省次、加藤剛二、加藤俊一、麦島秀雄、土田昌宏
移植実施調査期間
1985~ 2002年
初回移植症例数
123例再移植
15回、 3回目の移植 1回
実施施設
35施設
疾患ムコ多糖症: 63 ( I:9, IS:3, II:39, III:2, IV:4, VI:4, VII:2)副腎白質ジストロフィー: 35I-cell病: 6異染性ロイコジストロフィ: 5Gaucher病: 4GM1 gangliosidosis: 4
その他: 6( Krabbe, Galactosialidosis, Alfa-Mannosidosis, Nieman-Pick, Pompe, Multiple sulfatase def.)
性別 男性: 100, 女性: 23初回移植時年齢
0歳: 5, 1歳: 9, 1歳: 19, 3歳:10 、 4歳: 16, 5歳: 10, 6歳: 11,7歳: 12 、 8歳: 7, 9歳: 6, 10歳以上: 18
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移植の概要(初回移植)• ドナー 同胞:59例、両親:7例、非血縁者:57• HLA適合度 一致:98例、部分一致:24例、不明:1例• 移植細胞源 骨髄:103例、末梢血+骨髄:1例、 臍帯血:19例
移植の結果(初回移植)• 生着 あり: 81例 ( 65.8%) なし(拒絶): 33例 ( 26.8%) 不明: 9例 ( 7.3%)• 転帰 生存: 99例 ( 80.5%) 死亡: 24例 ( 19.5%)
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平成 15 年 平成 16 年 平成 17 年細胞の調整(骨髄由来細胞)・培養法の開発と改良
バリデーション(移植、 Chip 、核型、表面マーカー、分化形質)
倫理委員会審査中
疾病モデルマウスの SCID 化
間葉系細胞による「皮膚創傷治癒」に対する臨床研究
大動物(ブタ)による移植実験
ヒト細胞に関する CPC の設立 臨床プロトコールの作製・倫理委員会審査
国立成育医療センターにおけるCell Processing Center
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細胞を用いたムコ多糖症 VII 型マウスに対する中枢神経病変に対する細胞治療・遺伝子治療(大橋十也、島田隆)
出生前診断・胎児治療(末岡浩、林聡)
先天性代謝異常症に対する造血幹細胞移植日本国における現状(加藤俊一)
細胞治療におけるヒト細胞の安全性の検討ならびにバリデーション(梅澤明弘)
リソソーム病とは
遺伝的要因によるリソソーム酵素活性の低下
これがリソソーム外に排泄されず進行的に蓄積する。
細胞機能が低下し、臨床症状を呈する。
特異顔貌、
骨形成異常、
聴覚・視覚・心血管異常
関節可動域の異常、
精神発達遅滞
先天性代謝異常症に対する幹細胞治療法の開発
臨床研究に関する倫理指針厚生労働省、平成 16年 12月 28 日改訂
ヒト幹細胞等を用いる臨床研究に関する指針厚生労働省、未定稿
国立成育医療センター倫理委員会
ヒト幹細胞推進プロジェクト会議
細胞供給源骨髄由来細胞 (大橋、梅澤、加藤、宮本)脂肪細胞 (島田)唾液腺上皮 (中村)
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慶應義塾大学微生物学教室瀬川 薫
埼玉医科大学五條 理志許 俊英
血液内科木崎 昌弘
福田 恵一牧野 伸司
産婦人科吉村 泰典 丸山 哲也
循環器内科小川 聡三好 俊一郎西山 信大
国立がんセンター清野 透
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慶應義塾大学病理学教室森 泰昌竹田征治
循環器内科小川 聡福田恵一牧野伸司
生理学教室岡野栄之島崎琢也
埼玉医科大学五條理志許 俊英
慶應義塾大学血液内科池田 康夫木崎 昌弘
産婦人科吉村 泰典 丸山 哲也
整形外科学教室戸山芳昭今林英明越智健介
微生物学教室瀬川 薫
NIH, USA洪 実
熊本大学田賀哲也中嶋欽一
近畿大学角田幸雄加藤容子
National Cancer Center, Jpn清野 透
Helix研究所磯貝 隆夫杉山 友康入江 亮太郎
中外製薬東佐由美大川広行
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Takeda
Mori Tsuchiya
Matsumoto
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National Center for Child Health and Development