Nestin expression in mesenchymal stromal cells: Regulation by hypoxia and osteogenesis

Alice Wong, Ehssan Ghassemi, Clare E Yellowley-genetos

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Background: The intermediate filament protein nestin is used as a marker for neural stem cells, and its expression is inversely correlated with cellular differentiation. More recently, nestin expression has also been described in other cell types including multipotential mesenchymal stromal cells (MSCs). In this study, we examined the expression of nestin in equine, canine and human bone marrow-derived MSCs undergoing osteogenic differentiation, to determine whether nestin levels were attenuated as the cells acquired a more mature phenotype. In addition, the expression of nestin may be under the influence of cellular hypoxia, as nestin expression is known to increase in areas of ischemic tissue damage. Therefore, we also examined the effects of hypoxia on expression of nestin in human MSCs and examined a role for hypoxia inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor (VEGF) in the response. Additionally, we quantified the temporal expression of nestin in the fracture callus during bone regeneration, a site that has been characterized as hypoxic. Results: There were no significant changes in nestin expression in MSCs during osteogenic differentiation. There was a significant increase in expression of nestin mRNA and protein in human MSCs in response to hypoxia (1% O2) or the chemical hypoxia mimetic desferroxamine. This may be due to upregulation of VEGF under hypoxia, as treatment of cells with the VEGF receptor antagonist CPO-P11 attenuated hypoxia-induced nestin expression. A significant increase in nestin mRNA expression was observed in the fracture callus of mice three and seven days post fracture. Conclusions: Nestin was not a selective marker for MSCs, as its expression was maintained during osteogenic differentiation, in all species examined. Furthermore our data suggest that nestin expression can be induced by hypoxia, and that this increase in nestin is partially regulated by HIF-1α and VEGF. Interestingly, nestin levels were significantly upregulated at the fracture site. Further studies are required to understand the role of nestin in bone cell biology and ultimately bone regeneration.

Original languageEnglish (US)
Article number173
JournalBMC Veterinary Research
Volume10
Issue number1
DOIs
StatePublished - Aug 5 2014

Fingerprint

Cell Hypoxia
Nestin
stromal cells
bone formation
Mesenchymal Stromal Cells
Osteogenesis
hypoxia
vascular endothelial growth factors
bones
callus
intermediate filament proteins
vascular endothelial growth factor receptors
Vascular Endothelial Growth Factor A
Hypoxia-Inducible Factor 1
cells
Bone Regeneration
cell biology
bone marrow
Bony Callus
stem cells

Keywords

  • Canine
  • Equine
  • Fracture callus
  • Human
  • Mesenchymal stromal cells
  • Nestin
  • Osteogenesis
  • VEGF

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Nestin expression in mesenchymal stromal cells : Regulation by hypoxia and osteogenesis. / Wong, Alice; Ghassemi, Ehssan; Yellowley-genetos, Clare E.

In: BMC Veterinary Research, Vol. 10, No. 1, 173, 05.08.2014.

Research output: Contribution to journalArticle

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abstract = "Background: The intermediate filament protein nestin is used as a marker for neural stem cells, and its expression is inversely correlated with cellular differentiation. More recently, nestin expression has also been described in other cell types including multipotential mesenchymal stromal cells (MSCs). In this study, we examined the expression of nestin in equine, canine and human bone marrow-derived MSCs undergoing osteogenic differentiation, to determine whether nestin levels were attenuated as the cells acquired a more mature phenotype. In addition, the expression of nestin may be under the influence of cellular hypoxia, as nestin expression is known to increase in areas of ischemic tissue damage. Therefore, we also examined the effects of hypoxia on expression of nestin in human MSCs and examined a role for hypoxia inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor (VEGF) in the response. Additionally, we quantified the temporal expression of nestin in the fracture callus during bone regeneration, a site that has been characterized as hypoxic. Results: There were no significant changes in nestin expression in MSCs during osteogenic differentiation. There was a significant increase in expression of nestin mRNA and protein in human MSCs in response to hypoxia (1{\%} O2) or the chemical hypoxia mimetic desferroxamine. This may be due to upregulation of VEGF under hypoxia, as treatment of cells with the VEGF receptor antagonist CPO-P11 attenuated hypoxia-induced nestin expression. A significant increase in nestin mRNA expression was observed in the fracture callus of mice three and seven days post fracture. Conclusions: Nestin was not a selective marker for MSCs, as its expression was maintained during osteogenic differentiation, in all species examined. Furthermore our data suggest that nestin expression can be induced by hypoxia, and that this increase in nestin is partially regulated by HIF-1α and VEGF. Interestingly, nestin levels were significantly upregulated at the fracture site. Further studies are required to understand the role of nestin in bone cell biology and ultimately bone regeneration.",
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KW - Osteogenesis

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