Bone Matrix Formation in Osteogenic Cultures Derived from Human Embryonic Stem Cells In Vitro

Kärner, Elerin; Unger, Christian; Sloan, Alastair James; Ährlund‐Richter, Lars; Sugars, Rachael V.; Wendel, Mikael

doi:10.1089/scd.2007.16.ft-1

Cited by 15 publications

(25 citation statements)

References 50 publications

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“…In support of this notion, several studies have used the hEB model to differentiate hESCs into mesenchymal osteoblastic cells (22) and other cell types (neural-crest-like cells). (63) While some studies succeeded in differentiating hESCs directly from monolayer cultures (19,20) our study demonstrates some advantages of using the EB formation step.…”

Section: Discussionmentioning

confidence: 75%

“…(13) Recent publications have demonstrated the development of clinically suitable ex vivo differentiation protocols for obtaining insulin-producing b cells (14,15) and neuronal cells (16,17) from hESCs. Several protocols have been reported in which hESCs were directed to differentiate into musculoskeletal tissue: osteoblastic cells, (18)(19)(20)(21)(22) chondroyctic cells, (23) cardiomyocytes, (24)(25)(26)(27)(28) and skeletal myoblasts. (29) However, most of these protocols relied on using coculture with differentiated cells or complex culture conditions to induce differentiation.…”

Section: Introductionmentioning

confidence: 99%

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Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-β/activin/nodal signaling using SB-431542

Mahmood

Harkness²,

Schrøder

et al. 2010

Journal of Bone and Mineral Research

120

100

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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 b (TGF-b)/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), a-smooth muscle actin, Nkx2.5, cTNT]. Continuous inhibition of TGF-b signaling in EB outgrowth cultures (SB-OG) enriched for myocyte progenitor cells; markers were PAX7 þ (25%), MYOD1 þ (52%), and NCAM þ (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 þ (100%), CD73 þ (98%), CD146 þ (96%), and CD166 þ (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 (À167.9-fold), ACTA1 (À161-fold), MYBPH (À139-fold), ACTC (À100.3-fold), MYH8 (À45.5-fold), and MYOT (À41.8-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-b signaling pathway. ß

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Section: Discussionmentioning

confidence: 75%

Section: Introductionmentioning

confidence: 99%

Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-β/activin/nodal signaling using SB-431542

Mahmood

Harkness²,

Schrøder

et al. 2010

Journal of Bone and Mineral Research

120

100

View full text Add to dashboard Cite

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“…Our own studies have shown that HESC lines differentiate along the osteogenic lineage, forming a fully mineralized bone-like matrix (Kärner et al, 2009;Kärner et al, 2007). We demonstrated the osteoblast phenotype using a large panel of extracellular matrix molecules and transcription factors (Kärner et al, 2007), and showed the dynamic gene expression of these markers (Kärner et al, 2009). In addition we characterized the deposited mineral with Fourier InfraRed spectroscopy proving that it resembled natural bone and was formed by cell-mediated mineralization .…”

Section: Cellsmentioning

confidence: 70%

“…Many groups have performed direct osteogenic differentiation, whereas others have taken the cells through a MSC progenitor stage prior to osteoblasts (Arpornmaeklong et al, 2009;Brown et al, 2009;Karp et al, 2006). Our own studies have shown that HESC lines differentiate along the osteogenic lineage, forming a fully mineralized bone-like matrix (Kärner et al, 2009;Kärner et al, 2007). We demonstrated the osteoblast phenotype using a large panel of extracellular matrix molecules and transcription factors (Kärner et al, 2007), and showed the dynamic gene expression of these markers (Kärner et al, 2009).…”

Section: Cellsmentioning

confidence: 81%

Clinical Concepts in Oral and Maxillofacial Surgery and Novel Findings to the Field of Bone Regeneration

Rosén¹,

Sugars²

2012

Bone Grafting

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“…hESC differentiation to osteoblasts has been studied extensively [98], and crucial transcription factors regulating osteoblast differentiation have been identified together with their specific function [99][100][101][102][103] such as Runx2, Osterix, b-catenin, Foxc1, Msx1, Msx2, Dlx5, Dlx6, Twist, AP1(Fos/Jun), Knox-20, Sp3, Atf4, Alx4, etc. Among those supremacy has been awarded to the early osteogenic marker Runx2 and the late osteogenic regulator Osterix [104][105][106][107], with particularly Runx2 playing a master regulatory role and being indispensable for osteoblast differentiation [107].…”

Section: Reprogramming Factorsmentioning

confidence: 99%

Concise Review: Induced Pluripotent Stem Cells and Lineage Reprogramming: Prospects for Bone Regeneration

et al. 2011

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Bone tissue for transplantation therapies is in high demand in clinics. Osteodegenerative diseases, in particular, osteoporosis and osteoarthritis, represent serious public health issues affecting a respectable proportion of the elderly population. Furthermore, congenital indispositions from the spectrum of craniofacial malformations such as cleft palates and systemic disorders including osteogenesis imperfecta are further increasing the need for bone tissue. Additionally, the reconstruction of fractured bone elements after accidents and the consumption of bone parts during surgical tumor excisions represent frequent clinical situations with deficient availability of healthy bone tissue for therapeutic transplantations. Epigenetic reprogramming represents a powerful technology for the generation of healthy patientspecific cells to replace or repair diseased or damaged tissue. The recent generation of induced pluripotent stem cells (iPSCs) is probably the most promising among these approaches dominating the literature of current stem cell research. It allows the generation of pluripotent stem cells from adult human skin cells from which potentially all cell types of the human body could be obtained. Another technique to produce clinically interesting cell types is direct lineage reprogramming (LR) with the additional advantage that it can be applied directly in vivo to reconstitute a damaged organ. Here, we want to present the two technologies of iPSCs and LR, to outline the current states of research, and to discuss possible strategies for their implementation in bone regeneration.

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Bone Matrix Formation in Osteogenic Cultures Derived from Human Embryonic Stem Cells In Vitro

Cited by 15 publications

References 50 publications

Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-β/activin/nodal signaling using SB-431542

Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-β/activin/nodal signaling using SB-431542

Clinical Concepts in Oral and Maxillofacial Surgery and Novel Findings to the Field of Bone Regeneration

Concise Review: Induced Pluripotent Stem Cells and Lineage Reprogramming: Prospects for Bone Regeneration

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