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Stem cells and development2017; 26(6); 441-450; doi: 10.1089/scd.2016.0279

Scleraxis Is Essential for Tendon Differentiation by Equine Embryonic Stem Cells and in Equine Fetal Tenocytes.

Abstract: The transcription factor scleraxis is required for tendon development and is upregulated during embryonic stem cell (ESC) differentiation into tenocytes. However, its role beyond early embryonic development is not defined. We utilized a short hairpin RNA to knock down scleraxis expression in ESCs and adult and fetal tenocytes. No effect on growth or morphology was observed in two-dimensional cultures. However, scleraxis knockdown in fetal tenocytes significantly reduced COL1A1, COMP, and SOX9 gene expression. Scleraxis knockdown in adult tenocytes had no effect on the expression of these genes. Strikingly, differentiating ESCs and fetal tenocytes without scleraxis failed to reorganize a three-dimensional (3D) matrix and generate artificial tendons. This was associated with a significantly reduced survival. In contrast, there was no effect on the survival and remodeling capacity of adult tenocytes following scleraxis knockdown. Overexpression of scleraxis in fetal tenocytes rescued gene expression, cell survival in 3D, and subsequent matrix contraction. Together, these results demonstrate that scleraxis is not only essential for ESC differentiation into tenocytes but that it also has an active role in maintaining fetal tenocytes, which is then redundant in adult tenocytes.
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Publication Date: 2017-01-24 PubMed ID: 27899062DOI: 10.1089/scd.2016.0279Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research shows that a transcription factor named scleraxis, which is known to be necessary for tendon development and increases during embryonic stem cell differentiation into tenocytes, also plays an important role in maintaining fetal tendon cells. The knockdown of scleraxis results in the inability to generate artificial tendons, even though it does not impact the growth or shape of cells in standard cultures.

Objective, Design, and Methodology of the Research

  • The researchers used a technique known as short hairpin RNA (shRNA) to suppress the expression of the scleraxis transcription factor in embryonic stem cells (ESCs), and in adult and fetal tenocytes (tendon cells). This technique, known as silencing or ‘knockdown’, lets scientists explore the function of particular genes by reducing their activity.
  • The experiments were conducted in both two-dimensional cultures and three-dimensional (3D) matrices to evaluate the impact of scleraxis knockdown on cell behavior in both simple and complex environments. In the 3D matrix, the cells responded to the knockdown by reorganizing themselves and generating synthetic tendons.

Findings of the Research

  • Silencing scleraxis in ESCs and fetal tenocytes led to a significant reduction of specific gene expressions (COL1A1, COMP, and SOX9), essential for tendon development. But, silencing scleraxis in adult tenocytes did not affect the expression of these genes.
  • Remarkably, ESCs and fetal tenocytes without scleraxis were incapable of reorganizing in a 3D matrix and creating artificial tendons. They also exhibited a significantly reduced survival rate. However, adult tenocytes didn’t show any change in their survival and remodeling capacity post the scleraxis knockdown.
  • The researchers were able to reverse the effects of scleraxis suppression in fetal tenocytes by reintroducing scleraxis. This ‘rescue’ restored gene expression, cell survival in 3D, and subsequent matrix contraction.

Implications of the Research

  • The study thus implies that the scleraxis transcription factor is not only crucial for the differentiation of ESCs into tenocytes (tendon cells) but also plays a key role in maintaining fetal tenocytes.
  • Scleraxis’s role seems to become redundant in adult tenocytes, who can maintain themselves and remodel their structure even when its expression is inhibited.

Cite This Article

APA
Bavin EP, Atkinson F, Barsby T, Guest DJ. (2017). Scleraxis Is Essential for Tendon Differentiation by Equine Embryonic Stem Cells and in Equine Fetal Tenocytes. Stem Cells Dev, 26(6), 441-450. https://doi.org/10.1089/scd.2016.0279

Publication

Stem cells and development
ISSN: 1557-8534
NlmUniqueID: 101197107
Country: United States
Language: English
Volume: 26
Issue: 6
Pages: 441-450

Researcher Affiliations

Bavin, Emma P
  • 1 Centre for Preventive Medicine , Animal Health Trust, Newmarket, United Kingdom .
Atkinson, Francesca
  • 1 Centre for Preventive Medicine , Animal Health Trust, Newmarket, United Kingdom .
Barsby, Tom
  • 1 Centre for Preventive Medicine , Animal Health Trust, Newmarket, United Kingdom .
Guest, Debbie J
  • 1 Centre for Preventive Medicine , Animal Health Trust, Newmarket, United Kingdom .

MeSH Terms

  • Aging / physiology
  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cartilage Oligomeric Matrix Protein / metabolism
  • Cell Differentiation / drug effects
  • Cell Differentiation / genetics
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • Collagen Type I / genetics
  • Collagen Type I / metabolism
  • DNA, Complementary / metabolism
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / drug effects
  • Embryonic Stem Cells / metabolism
  • Fetus / cytology
  • Gels / pharmacology
  • Gene Expression Regulation / drug effects
  • Gene Knockdown Techniques
  • Horses
  • RNA, Small Interfering / metabolism
  • SOX9 Transcription Factor / genetics
  • SOX9 Transcription Factor / metabolism
  • Tendons / cytology
  • Tenocytes / cytology
  • Tenocytes / drug effects
  • Tenocytes / metabolism

Citations

This article has been cited 16 times.
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