Characterization of extracellular matrix macromolecules in primary cultures of equine keratinocytes

Visser, Michelle B. and Pollitt, Christopher C. (2010) Characterization of extracellular matrix macromolecules in primary cultures of equine keratinocytes. BMC Veterinary Research, 6 1-8. doi:10.1186/1746-6148-6-16


Author Visser, Michelle B.
Pollitt, Christopher C.
Title Characterization of extracellular matrix macromolecules in primary cultures of equine keratinocytes
Journal name BMC Veterinary Research   Check publisher's open access policy
ISSN 1746-6148
Publication date 2010-03-15
Sub-type Article (original research)
DOI 10.1186/1746-6148-6-16
Open Access Status DOI
Volume 6
Start page 1
End page 8
Total pages 8
Place of publication London, U.K.
Publisher BioMed Central
Collection year 2011
Language eng
Formatted abstract
Background: 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 (α6) and the hemidesmsome 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.
© 2010 Visser and Pollitt; licensee BioMed Central Ltd.

Keyword Human epidermal-keratinocytes
Basement-membrane
In-vitro
Cells
Hemidesmosomes
Cultivation
Laminin-5
Protein
Growth
Differentiation
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published online only - Article number 16

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2011 Collection
School of Veterinary Science Publications
 
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Citation counts: TR Web of Science Citation Count  Cited 8 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 9 times in Scopus Article | Citations
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Created: Sun, 25 Apr 2010, 00:06:13 EST