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American journal of veterinary research2000; 61(9); 1003-1010; doi: 10.2460/ajvr.2000.61.1003

Effect of transforming growth factor beta1 on chondrogenic differentiation of cultured equine mesenchymal stem cells.

Abstract: To determine the morphologic and phenotypic effects of transforming growth factor beta1 (TGFbeta1) on cultured equine mesenchymal stem cells (MSC) and articular chondrocytes. Methods: Bone marrow aspirates and articular cartilage samples from a 2-year-old and two 8-month-old horses. Methods: After initial isolation and culture, MSC and chondrocytes were cultured in Ham's F-12 medium supplemented with TGF-beta1 at a concentration of 0, 1, 5, or 10 ng/ml. Medium was exchanged on day 2, and cells were harvested on day 4. Medium was assayed for proteoglycan (PG) content. Total RNA was isolated from cell cultures, and expression of aggrecan, decrin, collagen type-I, and collagen type-II mRNA was assessed by means of Northern blot analyses. Cell cultures were stained with H&E or toluidine blue and examined histologically. Additional cultures were examined after immunohistochemical staining for type-I and -II collagen. Results: MSC cultures exposed to TGF-beta1 had an increased cellular density with cell layering and nodule formation that was most pronounced in cultures treated with 5 ng of TGF-beta1/ml. Expression of collagen type-II mRNA in MSC cultures exposed to 5 ng of TGF-beta1/ml was 1.7 times expression in control cultures, and expression of collagen type-I mRNA was 2.8 times expression in control cultures. Treatment of MSC with TGF-beta1 led to dose-related increases in area and intensity of type-II collagen immunoreaction. Conclusions: Results suggest that TGF-beta1 enhances chondrogenic differentiation of bone marrow-derived MSC in a dose-dependent manner.
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Publication Date: 2000-09-08 PubMed ID: 10976727DOI: 10.2460/ajvr.2000.61.1003Google 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.

The research focused on examining the influence of transforming growth factor beta1 (TGF-beta1) on equine mesenchymal stem cells (MSC) and their differentiation into cartilage cells. The key finding is that TGF-beta1 promotes this differentiation in higher amounts, evident from an increased cellular density, collagen type-II mRNA expression, and enhanced type-II collagen immunoreaction.

Introduction and Methodology

  • The research was conducted utilizing bone marrow aspirates (sources of MSCs) and articular cartilage samples from several horses, which were isolated and cultured.
  • Post-isolation, the MSCs and cartilage cells (chondrocytes) were then exposed to TGF-beta1, a protein that regulates cellular functions, at varying concentrations to observe its effect.
  • The researchers studied the impact of TGF-beta1 by analysing the cell’s response via changes in morphology and gene expression, measured by staining techniques, analysis of cellular components like proteoglycans (PG), and performing Northern blot analyses for mRNA expression of various proteins.

Results

  • Upon exposure to TGF-beta1, MSC cultures showed an increased cellular density, indicating potential growth or proliferation.
  • There was notable cell layering and nodule formation, especially in cultures treated with 5ng of TGF-beta1 for each milliliter, suggesting that a specific concentration of TGF-beta1 drives the most significant change.
  • In terms of gene expression, there was an increase in the expression of collagen type-II mRNA, nearly 1.7 times more than in control cultures. This is indicative of enhanced chondrogenic differentiation as collagen type-II is a principal component of cartilage.
  • Expression levels of collagen type-I mRNA were also observed to have increased, by 2.8 times, compared to the control cultures.
  • Through immunohistochemical staining, an increase in the area and intensity of type-II collagen reactions was noted, suggesting an increased differentiation and maturation of MSCs into chondrocytes.

Conclusion

  • The research concludes that TGF-beta1 plays a vital role in promoting chondrogenic differentiation of equine MSCs. The process is dose-dependent, with specific concentrations showing a more prominent effect.
  • The findings can provide a foundation for therapeutic strategies, particularly in regenerative medicine, by controlling and enhancing the differentiation of stem cells into required cell types through manipulation of growth factors like TGF-beta1.

Cite This Article

APA
Worster AA, Nixon AJ, Brower-Toland BD, Williams J. (2000). Effect of transforming growth factor beta1 on chondrogenic differentiation of cultured equine mesenchymal stem cells. Am J Vet Res, 61(9), 1003-1010. https://doi.org/10.2460/ajvr.2000.61.1003

Publication

American journal of veterinary research
ISSN: 0002-9645
NlmUniqueID: 0375011
Country: United States
Language: English
Volume: 61
Issue: 9
Pages: 1003-1010

Researcher Affiliations

Worster, A A
  • Comparative Orthopaedics Laboratory, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Nixon, A J
    Brower-Toland, B D
      Williams, J

        MeSH Terms

        • Animals
        • Blotting, Northern / veterinary
        • Bone Marrow Cells / cytology
        • Cartilage, Articular / cytology
        • Cartilage, Articular / drug effects
        • Cell Differentiation / drug effects
        • Cells, Cultured
        • Dose-Response Relationship, Drug
        • Horses
        • In Situ Hybridization
        • Phenotype
        • Stem Cells / drug effects
        • Transforming Growth Factor beta / pharmacology

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