Transcriptional signature of human adipose tissue-derived stem cells (hASCs) preconditioned for chondrogenesis in hypoxic conditions

Pilgaard, Linda; Lund, Pia; Duroux, Meg; Lockstone, Helen; Taylor, John; Emmersen, Jeppe; Fink, Trine; Ragoussis, Jiannis; Zachar, Vladimír

doi:10.1016/j.yexcr.2009.01.020

Cited by 49 publications

(38 citation statements)

References 95 publications

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“…Furthermore, hypoxia provides a safe, clinically compliant alternative to other methods, such as genetic modification, which could be used to protect stem cells in harsh ischemic environments. Long-term hypoxia has been shown to affect the proliferation and differentiation of ASC [26][27][28], but the cytoprotective effects of HPC have not been properly investigated. Ours is the first study to demonstrate clearly that HPC can protect ASC against ischemic injury in vitro, and here we evaluate the mechanisms involved.…”

Section: Discussionmentioning

confidence: 99%

Hypoxic Preconditioning Enhances Survival of Human Adipose-Derived Stem Cells and Conditions Endothelial Cells In Vitro

Stubbs

Hsiao²,

Peshavariya³

et al. 2012

Stem Cells and Development

112

100

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To grow more robust cardiac tissue for implantation in vivo, strategies to improve survival of implanted stem cells are required. Here we report the protective effects of hypoxic preconditioning (HPC) and identify mechanisms for improving survival of adipose-derived stem cells (ASC) in vitro. Human ASC were preconditioned for 24 h with hypoxia and then exposed to simulated ischemia for a further 24 h. HPC significantly increased ASC viability, and reduced cell injury and apoptosis compared with non-preconditioned cells under ischemic conditions, as shown by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), lactate dehydrogenase-release, and caspase activity assays. Preconditioned ASC increased levels of hypoxia-inducible factor-1 alpha and secreted significantly more of the downstream target vascular endothelial growth factor (VEGF-A; 13-fold) compared with control during the 24 h. Exogenous VEGF (50 ng/mL) increased phosphorylation of Akt without affecting ERK1/2, JNK, or p38 MAPK protein levels. Phospho-Akt was also increased in preconditioned ASC compared with non-preconditioned ASC, an effect that may be mediated via VEGF-A. Importantly, the protective effects of HPC were abolished by a neutralizing antibody against VEGF-A and the phosphoinositol 3-kinase inhibitor LY294002, demonstrating the importance of VEGF-A and Akt in hypoxia-induced ASC survival. Importantly, we showed that media derived from hypoxic preconditioned ASC support endothelial cell survival and endothelial tube formation in vitro. Our in vitro findings indicate that HPC may be a promising strategy to improve survival of ASC and promote angiogenesis in ischemic environments.

show abstract

Section: Discussionmentioning

confidence: 99%

Hypoxic Preconditioning Enhances Survival of Human Adipose-Derived Stem Cells and Conditions Endothelial Cells In Vitro

Stubbs

Hsiao²,

Peshavariya³

et al. 2012

Stem Cells and Development

112

100

View full text Add to dashboard Cite

show abstract

“…Expression of the facilitative glucose transporter GLUT1 (SLC2A1), and of genes encoding glycolytic enzymes are recognised to be hypoxia-sensitive (Pilgaard et al, 2009;Wood et al, 2007). As in our earlier candidate gene study, increased expression of GLUT1 and GLUT3 was found here on the microarrays (5.6-and 3.1-fold increase in mRNA level, respectively), whereas expression of the other glucose transporters, including the insulin-stimulated glucose transporter, GLUT4, was not altered by hypoxia.…”

Section: Discussionmentioning

confidence: 99%

A microarray analysis of the hypoxia-induced modulation of gene expression in human adipocytes

Mazzatti

Lim

O’Hara

et al. 2012

Archives of Physiology and Biochemistry

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“…Indeed, recent works have shown that leptin inhibits chondrogenesis [26]. Blaney Davidson et al showed an increased ALK-1/ALK-5 ratio as a cause of deviant chondrocyte behavior (elevated MMP-13 expression), which contributes to age-related cartilage destruction and osteoarthritis in humans [27,28].…”

Section: Discussionmentioning

confidence: 99%

Differential Signalling Through ALK-1 and ALK-5 Regulates Leptin Expression in Mesenchymal Stem Cells

Zeddou¹,

Relić

Malaise

et al. 2012

Stem Cells and Development

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Leptin plays a central role in maintaining energy balance, with multiple other systemic effects. Despite leptin importance in peripheral regulation of mesenchymal stem cells (MSC) differentiation, little is known about its expression mechanism. Leptin is often described as adipokine, while it is expressed by other cell types. We have recently shown an in vitro leptin expression, enhanced by glucocorticoids in synovial fibroblasts (SVF). Here, we investigated leptin expression in MSC from bone marrow (BM-MSC) and umbilical cord matrix (UMSC). Results showed that BM-MSC, but not UMSC, expressed leptin that was strongly enhanced by glucocorticoids. Transforming growth factor b1 (TGF-b1) markedly inhibited the endogenous-and glucocorticoid-induced leptin expression in BM-MSC. Since TGF-b1 was shown to signal via ALK-5-Smad2/3 and/or ALK-1-Smad1/5 pathways, we analyzed the expression of proteins from both pathways. In BM-MSC, TGF-b1 increased phosphorylated Smad2 (p-Smad2) expression, while ALK-5 inhibitor (SB431542) induced leptin expression and significantly restored TGF-b1-induced leptin inhibition. In addition, both prednisolone and SB431542 increased p-Smad1/5 expression. These results suggested the ALK-5-Smad2 pathway as an inhibitor of leptin expression, while ALK-1-Smad1/5 as an activator. Indeed, Smad1 expression silencing induced leptin expression inhibition. Furthermore, prednisolone enhanced the expression of TGF-bRII while decreasing p-Smad2 in BM-MSC and SVF but not in UMSC. In vitro differentiation revealed differential osteogenic potential in SVF, BM-MSC, and UMSC that was correlated to their leptin expression potential. Our results suggest that ALK-1/ALK-5 balance regulates leptin expression in MSC. It also underlines UMSC as leptin nonproducer MSC for cell therapy protocols where leptin expression is not suitable.

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Transcriptional signature of human adipose tissue-derived stem cells (hASCs) preconditioned for chondrogenesis in hypoxic conditions

Cited by 49 publications

References 95 publications

Hypoxic Preconditioning Enhances Survival of Human Adipose-Derived Stem Cells and Conditions Endothelial Cells In Vitro

Hypoxic Preconditioning Enhances Survival of Human Adipose-Derived Stem Cells and Conditions Endothelial Cells In Vitro

A microarray analysis of the hypoxia-induced modulation of gene expression in human adipocytes

Differential Signalling Through ALK-1 and ALK-5 Regulates Leptin Expression in Mesenchymal Stem Cells

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