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Home / Equine Research Bank / BMC Vet Res / Characterization of extracellular matrix macromolecules in p...
BMC veterinary research2010; 6; 16; doi: 10.1186/1746-6148-6-16

Characterization of extracellular matrix macromolecules in primary cultures of equine keratinocytes.

Abstract: Most research to date involving laminins and extracellular matrix protein function in both normal and pathological conditions involves in vitro culture of keratinocytes. Few methods are established to allow for prolonged propagation of keratinocytes from equine tissues, including the hoof lamellae. In this study we modified cell isolation and culture techniques to allow for proliferation and sub-culturing of equine lamellar keratinocytes. Additionally, the production and processing of extracellular matrix molecules by skin and lamellar keratinocytes were studied. Results: Physical and proteolytic tissue separation in combination with media containing a calcium concentration of 0.6 mM in combination with additional media supplements proved optimal for proliferation and subculture of equine lamellar keratinocytes on collagen coated substratum. Immunofluorescence and immunoblotting studies confirmed that equine skin and lamellar keratinocytes produce Ln-332 in vitro and processing of this molecule follows that of other species. As well, matrix components including integrin alpha-6 (alpha 6) and the hemidesmosome proteins, bullous pemphigoid antigen 1 (BP180) bullous pemphigoid antigen 2 (BP230) and plectin are also expressed. Conclusions: Isolation of equine keratinocytes and study of the matrix and adhesion related molecules produced by them provides a valuable tool for future work in the veterinary field.
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Publication Date: 2010-03-15 PubMed ID: 20230631PubMed Central: PMC2847556DOI: 10.1186/1746-6148-6-16Google Scholar: Lookup
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  • 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 focuses on modifying cell isolation and culture techniques to allow for the growth and development of equine keratinocytes, cells from horse tissues, specifically hoof lamellae. It examines the production and processing of proteins in the extracellular matrix, which is crucial for cell communication and function, in both skin and hoof keratinocytes.

Cell Isolation and Culture Modifications

  • The primary concern of the study was to find an appropriate method for breeding equine keratinocytes, particularly those from the hoof lamella. Traditional methods are limited and do not accommodate long-term propagation of the cells.
  • In order to optimize conditions for the propagation and culture of these cells, this study used a combination of physical and proteolytic separation techniques. These methods involve chemical and physical means to segregate the cells from the tissues.
  • This, coupled with media that had a calcium concentration of 0.6 mM and added media supplements, was found to be the best solution for the growth and sub-culture of equine lamellar keratinocytes on collagen-coated substratum.

Extracellular Matrix Molecule Production

  • The researchers then turned their attention to the extracellular matrix molecules produced by the skin and hoof keratinocytes. The extracellular matrix is essential for cell function, providing structural and biochemical support to surrounding cells.
  • Immunofluorescence and immunoblotting studies, which involve labelling antibodies to see where they bind in the tissue, revealed that both types of keratinocytes produce Ln-332 in vitro. Ln-332 is a type of laminin, which is critical for the structure and function of the matrix.
  • The research showed that the processing of the Ln-332 molecule aligns with that of other species, providing a generalizable outcome.

Expression of Matrix Components

  • Further studies showed that the matrix components integrin alpha-6 (alpha 6), and the hemidesmosome proteins bullous pemphigoid antigen 1 (BP180), bullous pemphigoid antigen 2 (BP230) and plectin are also expressed by equine keratinocytes.
  • These molecules are important for maintaining cell structure and facilitating cell-extracellular matrix adhesion, essential for cellular stability and functionality.

Conclusion

  • Ultimately the study contributes valuable tools and understanding to veterinary research. By establishing a consistently successful isolation and culture method for equine keratinocytes, and demonstrating the extracellular matrix-related molecules they produce, future research in the veterinary field can be expanded.

Cite This Article

APA
Visser MB, Pollitt CC. (2010). Characterization of extracellular matrix macromolecules in primary cultures of equine keratinocytes. BMC Vet Res, 6, 16. https://doi.org/10.1186/1746-6148-6-16

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 6
Pages: 16

Researcher Affiliations

Visser, Michelle B
  • The Australian Equine Laminitis Research Unit, School of Veterinary Science, University of Queensland, St Lucia, 4072, Australia. michelle.visser@utoronto.ca
Pollitt, Christopher C

    MeSH Terms

    • Animals
    • Cell Culture Techniques / veterinary
    • Cells, Cultured
    • Extracellular Matrix Proteins / metabolism
    • Gene Expression Regulation
    • Hemidesmosomes / metabolism
    • Keratinocytes / cytology
    • Keratinocytes / metabolism

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    Citations

    This article has been cited 6 times.
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