Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-beta/Activin/Nodal signaling using SB-431542

Amer Mahmood, Linda Harkness, Henrik Daa Schrøder, Basem Abdallah, Moustapha Kassem

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Udgivelsesdato: 2010-Jan-29
OriginalsprogEngelsk
TidsskriftJournal of Bone and Mineral Research
Vol/bind25
Udgave nummer6
Sider (fra-til)1216-1223
ISSN0884-0431
DOI
StatusUdgivet - 29. jan. 2010

Fingeraftryk

Inhibin-beta Subunits
Mesenchymal Stromal Cells
Up-Regulation
Regenerative Medicine
Desmin
Muscle Development
Serum-Free Culture Media
Cell Lineage
Mesoderm
Chondrocytes
Osteoblasts
Adipocytes
Population
Smooth Muscle
Actins
Cultured Cells
Down-Regulation
Human Embryonic Stem Cells
4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide
Serum

Citer dette

@article{3321b74037ff11df937d000ea68e967b,
title = "Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-beta/Activin/Nodal signaling using SB-431542",
abstract = "Directing differentiation of human embryonic stem cells (hESC) into specific cell types using an easy and reproducible protocol is a prerequisite for the clinical use of hESC in regenerative medicine procedures. Here, we report a protocol for directing the differentiation of hESC into mesenchymal progenitor cells. We demonstrate that inhibition of TGF-beta/Activin/Nodal signaling during embryoid bodies (EB) formation using SB-431542 (SB) in serum free medium, markedly up-regulated paraxial mesodermal markers (TBX6, TBX5), and several myogenic developmental markers including early myogenic transcriptional factors (Myf5, Pax7) as well as myocyte committed markers (NCAM, CD34, Desmin, MHC (fast), alpha-smooth muscle actin, Nkx2.5, cTNT). Continuous inhibition of TGF-beta signaling in EB outgrowth cultures (SB-OG) enriched for myocyte progenitor cells that were PAX7(+) (25{\%}), MYOD1(+) (52{\%}) and NCAM(+) (CD56) (73{\%}). DNA microarray analysis revealed differential up-regulation of 117 genes (>2-fold compared to control cells) annotated to myogenic development and function. Moreover, these cells showed the ability to contract (80{\%} of the population) and formed myofibres when implanted intramuscularly in vivo. Interestingly, SB-OG cells cultured in 10{\%} fetal bovine serum (FBS) developed into a homogeneous population of mesenchymal progenitors that expressed CD markers characteristic of mesenchymal stem cells (MSC): CD44(+) (100{\%}), CD73(+) (98{\%}), CD146(+) (96{\%}) and CD166(+) (88{\%}) with the ability to differentiate into osteoblasts, adipocytes and chondrocytes in vitro and in vivo. Furthermore, microarray analysis of these cells revealed down-regulation of genes related to myogenesis: MYH3 (-167.9 fold), ACTA1 (-161), MYBPH (-139 fold), ACTC (-100.3), MYH8 (-45.5 fold) and MYOT (-41.8 fold) and marked up-regulation of genes related to mesoderm-derived cell lineages. In conclusion, our data provides a simple and versatile protocol for directing the differentiation of hESC into a myogenic lineage and then further into mesenchymal progenitors by blocking the TGF-beta signaling pathway. (c) 2010 American Society for Bone and Mineral Research.",
author = "Amer Mahmood and Linda Harkness and Schr{\o}der, {Henrik Daa} and Basem Abdallah and Moustapha Kassem",
year = "2010",
month = "1",
day = "29",
doi = "10.1002/jbmr.34",
language = "English",
volume = "25",
pages = "1216--1223",
journal = "Journal of Bone and Mineral Research",
issn = "0884-0431",
publisher = "Wiley-Blackwell",
number = "6",

}

Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-beta/Activin/Nodal signaling using SB-431542. / Mahmood, Amer; Harkness, Linda; Schrøder, Henrik Daa; Abdallah, Basem; Kassem, Moustapha.

I: Journal of Bone and Mineral Research, Bind 25, Nr. 6, 29.01.2010, s. 1216-1223.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-beta/Activin/Nodal signaling using SB-431542

AU - Mahmood, Amer

AU - Harkness, Linda

AU - Schrøder, Henrik Daa

AU - Abdallah, Basem

AU - Kassem, Moustapha

PY - 2010/1/29

Y1 - 2010/1/29

N2 - Directing differentiation of human embryonic stem cells (hESC) into specific cell types using an easy and reproducible protocol is a prerequisite for the clinical use of hESC in regenerative medicine procedures. Here, we report a protocol for directing the differentiation of hESC into mesenchymal progenitor cells. We demonstrate that inhibition of TGF-beta/Activin/Nodal signaling during embryoid bodies (EB) formation using SB-431542 (SB) in serum free medium, markedly up-regulated paraxial mesodermal markers (TBX6, TBX5), and several myogenic developmental markers including early myogenic transcriptional factors (Myf5, Pax7) as well as myocyte committed markers (NCAM, CD34, Desmin, MHC (fast), alpha-smooth muscle actin, Nkx2.5, cTNT). Continuous inhibition of TGF-beta signaling in EB outgrowth cultures (SB-OG) enriched for myocyte progenitor cells that were PAX7(+) (25%), MYOD1(+) (52%) and NCAM(+) (CD56) (73%). DNA microarray analysis revealed differential up-regulation of 117 genes (>2-fold compared to control cells) annotated to myogenic development and function. Moreover, these cells showed the ability to contract (80% of the population) and formed myofibres when implanted intramuscularly in vivo. Interestingly, SB-OG cells cultured in 10% fetal bovine serum (FBS) developed into a homogeneous population of mesenchymal progenitors that expressed CD markers characteristic of mesenchymal stem cells (MSC): CD44(+) (100%), CD73(+) (98%), CD146(+) (96%) and CD166(+) (88%) with the ability to differentiate into osteoblasts, adipocytes and chondrocytes in vitro and in vivo. Furthermore, microarray analysis of these cells revealed down-regulation of genes related to myogenesis: MYH3 (-167.9 fold), ACTA1 (-161), MYBPH (-139 fold), ACTC (-100.3), MYH8 (-45.5 fold) and MYOT (-41.8 fold) and marked up-regulation of genes related to mesoderm-derived cell lineages. In conclusion, our data provides a simple and versatile protocol for directing the differentiation of hESC into a myogenic lineage and then further into mesenchymal progenitors by blocking the TGF-beta signaling pathway. (c) 2010 American Society for Bone and Mineral Research.

AB - Directing differentiation of human embryonic stem cells (hESC) into specific cell types using an easy and reproducible protocol is a prerequisite for the clinical use of hESC in regenerative medicine procedures. Here, we report a protocol for directing the differentiation of hESC into mesenchymal progenitor cells. We demonstrate that inhibition of TGF-beta/Activin/Nodal signaling during embryoid bodies (EB) formation using SB-431542 (SB) in serum free medium, markedly up-regulated paraxial mesodermal markers (TBX6, TBX5), and several myogenic developmental markers including early myogenic transcriptional factors (Myf5, Pax7) as well as myocyte committed markers (NCAM, CD34, Desmin, MHC (fast), alpha-smooth muscle actin, Nkx2.5, cTNT). Continuous inhibition of TGF-beta signaling in EB outgrowth cultures (SB-OG) enriched for myocyte progenitor cells that were PAX7(+) (25%), MYOD1(+) (52%) and NCAM(+) (CD56) (73%). DNA microarray analysis revealed differential up-regulation of 117 genes (>2-fold compared to control cells) annotated to myogenic development and function. Moreover, these cells showed the ability to contract (80% of the population) and formed myofibres when implanted intramuscularly in vivo. Interestingly, SB-OG cells cultured in 10% fetal bovine serum (FBS) developed into a homogeneous population of mesenchymal progenitors that expressed CD markers characteristic of mesenchymal stem cells (MSC): CD44(+) (100%), CD73(+) (98%), CD146(+) (96%) and CD166(+) (88%) with the ability to differentiate into osteoblasts, adipocytes and chondrocytes in vitro and in vivo. Furthermore, microarray analysis of these cells revealed down-regulation of genes related to myogenesis: MYH3 (-167.9 fold), ACTA1 (-161), MYBPH (-139 fold), ACTC (-100.3), MYH8 (-45.5 fold) and MYOT (-41.8 fold) and marked up-regulation of genes related to mesoderm-derived cell lineages. In conclusion, our data provides a simple and versatile protocol for directing the differentiation of hESC into a myogenic lineage and then further into mesenchymal progenitors by blocking the TGF-beta signaling pathway. (c) 2010 American Society for Bone and Mineral Research.

U2 - 10.1002/jbmr.34

DO - 10.1002/jbmr.34

M3 - Journal article

C2 - 20200949

VL - 25

SP - 1216

EP - 1223

JO - Journal of Bone and Mineral Research

JF - Journal of Bone and Mineral Research

SN - 0884-0431

IS - 6

ER -