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American journal of veterinary research2007; 68(9); 941-945; doi: 10.2460/ajvr.68.9.941

Effect of fibroblast growth factor-2 on equine mesenchymal stem cell monolayer expansion and chondrogenesis.

Abstract: To determine whether fibroblast growth factor-2 (FGF-2) treatment of equine mesenchymal stem cells (MSCs) during monolayer expansion enhances subsequent chondrogenesis in a 3-dimensional culture system. Methods: 6 healthy horses, 6 months to 5 years of age. Methods: Bone marrow-derived MSCs were obtained from 6 horses. First-passage MSCs were seeded as monolayers at 10,000 cells/cm(2) and in medium containing 0, 1, 10, or 100 ng of FGF-2/mL. After 6 days, MSCs were transferred to pellet cultures (200,000 cells/pellet) and maintained in chondrogenic medium. Pellets were collected after 15 days. Pellets were analyzed for collagen type II content by use of an ELISA, total glycosaminoglycan content by use of the dimethylmethylene blue dye-binding assay, and DNA content by use of fluorometric quantification. Semiquantitative PCR assay was performed to assess relative concentrations of collagen type II and aggrecan mRNAs. Results: Use of 100 ng of FGF-2/mL significantly increased pellet DNA and glycosaminoglycan content. Collagen type II content of the pellet was also increased by use of 10 and 100 ng of FGF-2/mL. Collagen type II and aggrecan mRNA transcripts were increased by treatment with FGF-2. Some control samples had minimal evidence of collagen type II and aggrecan transcripts after 35 cycles of amplification. Conclusions: FGF-2 treatment of bone marrow-derived MSC monolayers enhanced subsequent chondrogenic differentiation in a 3-dimensional culture. This result is important for tissue engineering strategies dependent on MSC expansion for cartilage repair.
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Publication Date: 2007-09-04 PubMed ID: 17764407DOI: 10.2460/ajvr.68.9.941Google 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 investigates the influence of a protein called fibroblast growth factor-2 (FGF-2) on the growth and function of horse stem cells, with a particular focus on its impact on the development of cartilage-like tissues. The findings of the study suggest that FGF-2 enhances the stem cells’ potential for cartilage development, which could be useful for therapeutic strategies involving cartilage repair.

Research Methodology

  • The researchers conducted this study on mesenchymal stem cells (MSCs) obtained from the bone marrow of six healthy horses aged between six months and five years.
  • The MSCs were grown as monolayers at a specific density and treated with varying concentrations of FGF-2 protein.
  • After six days, the MSCs were transferred to what’s referred to as ‘pellet cultures’. These are small, compact cell masses that are often used in studies of chondrogenesis, the formation of cartilage.
  • On the fifteenth day, these pellet cultures were collected and analyzed for various components related to cartilage formation, such as type II collagen, total glycosaminoglycan content, and DNA content. They also examined the presence of type II collagen and aggrecan mRNA transcripts—an indication of gene expression related to cartilage synthesis.

Research Findings

  • The results showed that the use of 100 ng of FGF-2/mL substantially increased the DNA content of the pellet and the glycosaminoglycan content. Glycosaminoglycans are a type of complex carbohydrate that plays a vital role in cartilage structure.
  • Increased levels of type II collagen, a major constituent of healthy cartilage, were observed in pellets that had been treated with 10 and 100 ng of FGF-2/mL.
  • Similarly, the presence of type II collagen and aggrecan mRNA transcripts were increased by treatment with FGF-2, indicating that the gene expression associated with cartilage formation was enhanced.

Conclusion

  • The research concluded that treating bone marrow-derived MSC monolayers with FGF-2 improved their potential for chondrogenic differentiation in a three-dimensional culture system.
  • These findings have significant implications for the field of tissue engineering, especially for strategies that rely on MSC expansion for the repair of damaged cartilage.

Cite This Article

APA
Stewart AA, Byron CR, Pondenis H, Stewart MC. (2007). Effect of fibroblast growth factor-2 on equine mesenchymal stem cell monolayer expansion and chondrogenesis. Am J Vet Res, 68(9), 941-945. https://doi.org/10.2460/ajvr.68.9.941

Publication

American journal of veterinary research
ISSN: 0002-9645
NlmUniqueID: 0375011
Country: United States
Language: English
Volume: 68
Issue: 9
Pages: 941-945

Researcher Affiliations

Stewart, Allison A
  • Department of Clinical Science, College of Veterinary Medicine, University of Illinois, Urbana, IL 61802, USA.
Byron, Christopher R
    Pondenis, Holly
      Stewart, Matthew C

        MeSH Terms

        • Aggrecans / biosynthesis
        • Aggrecans / genetics
        • Animals
        • Bisbenzimidazole / chemistry
        • Cell Culture Techniques
        • Cell Differentiation / drug effects
        • Collagen Type II / biosynthesis
        • Collagen Type II / genetics
        • DNA / analysis
        • Fibroblast Growth Factor 2 / pharmacology
        • Glycosaminoglycans / biosynthesis
        • Glycosaminoglycans / genetics
        • Histocytochemistry
        • Horses / physiology
        • Mesenchymal Stem Cells / cytology
        • Mesenchymal Stem Cells / drug effects
        • Mesenchymal Stem Cells / metabolism
        • Microscopy, Fluorescence / veterinary
        • RNA, Messenger / biosynthesis
        • RNA, Messenger / genetics
        • Reverse Transcriptase Polymerase Chain Reaction / veterinary
        • Tolonium Chloride / chemistry

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