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- W2023748814 abstract "Directing differentiation of human embryonic stem cells (hESCs) into specific cell types using an easy and reproducible protocol is a prerequisite for the clinical use of hESCs in regenerative-medicine procedures. Here, we report a protocol for directing the differentiation of hESCs into mesenchymal progenitor cells. We demonstrate that inhibition of transforming growth factor β (TGF-β)/activin/nodal signaling during embryoid body (EB) formation using SB-431542 (SB) in serum-free medium markedly upregulated 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), α-smooth muscle actin, Nkx2.5, cTNT]. Continuous inhibition of TGF-β signaling in EB outgrowth cultures (SB-OG) enriched for myocyte progenitor cells; markers were PAX7+ (25%), MYOD1+ (52%), and NCAM+ (CD56) (73%). DNA microarray analysis revealed differential upregulation of 117 genes (>2-fold compared with control cells) annotated to myogenic development and function. Moreover, these cells showed the ability to contract (80% of the population) and formed myofibers 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 (MSCs): 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 downregulation of genes related to myogenesis: MYH3 (−167.9-fold), ACTA1 (−161-fold), MYBPH (−139-fold), ACTC (−100.3-fold), MYH8 (−45.5-fold), and MYOT (−41.8-fold) and marked upregulation of genes related to mesoderm-derived cell lineages. In conclusion, our data provides a simple and versatile protocol for directing the differentiation of hESCs into a myogenic lineage and then further into mesenchymal progenitors by blocking the TGF-β signaling pathway. © 2010 American Society for Bone and Mineral Research" @default.
- W2023748814 created "2016-06-24" @default.
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- W2023748814 date "2010-01-29" @default.
- W2023748814 modified "2023-10-15" @default.
- W2023748814 title "Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-β/activin/nodal signaling using SB-431542" @default.
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- W2023748814 doi "https://doi.org/10.1002/jbmr.34" @default.
- W2023748814 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/20200949" @default.
- W2023748814 hasPublicationYear "2010" @default.
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