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Cultivation and characterisation of primary and subcultured equine keratinocytes.
Abstract: We describe the establishment and characterisation of equine keratinocyte cultures with maintenance of a high proliferative capacity up to the second passage. Improved attachment and growth were obtained by seeding primary cells on equine feeder layers. Subcultured keratinocytes showed optimal growth when seeded on collagen type I. The proliferation rate of cells on this substrate exceeded that seen for cells seeded on equine feeder layers. By immunohistochemistry, epithelial origin and state of differentiation of the equine keratinocytes were determined. They expressed keratin and desmoplakin I/II, but lacked keratin 10. Electron microscopy revealed typical features of cultured keratinocytes. Purity of keratinocyte cultures was determined by vimentin staining. This is the first report on the establishment of equine keratinocytes derived from lip epithelium. It forms the basis to study equine keratinocyte biology and the pathogenesis of epidermal diseases. Since wound healing represents a severe problem in equine dermatology, our data may be essential for the establishment of new and improved therapy.
Publication Date: 2002-03-21 PubMed ID: 11902754DOI: 10.2746/042516402776767187Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research discusses the development and analysis of equine keratinocyte cultures sustained with a high proliferative capacity up to their second passage. These are valuable when studying the biology of equine keratinocytes and the origin of skin disorders. The study also contributes to solving wound healing issues in equine dermatology.
Methodology
- The team created and detailed equine keratinocyte cultures. These are cells that produce keratin, a protein essential for the protection and repair of skin tissues.
- The cultures were maintained in a high proliferation capacity up to the second passage, meaning that they were capable of rapid growth and replication for extended periods.
- Improved attachment and growth were achieved by placing primary cells on equine feeder layers, suggesting that these cells perform best in their natural environment.
- Subcultured keratinocytes, cells propagated under lab conditions, showed optimal growth when seeded on collagen type I, outperforming those seeded on equine feeder layers in terms of their proliferation rate.
Findings
- Through immunohistochemistry (a technique used to visually see and localize antigens in cells of tissue), the origins and state of differentiation of the equine keratinocytes were determined.
- These cultured cells expressed keratin and desmoplakin I/II (key molecules in cell adhesion), but lacked keratin 10, showing that they were of a specific genetic makeup suitable for specific investigations into skin health.
- Observation through electron microscopy revealed ordinary features of cultured keratinocytes, which suggests that these cells retained their normal characteristics even in a lab environment.
- The purity of keratinocyte cultures was determined by vimentin staining. Vimentin is a type of intermediate filament (IF) protein that is often used as a marker of mesenchymal (connective tissue) cells.
Significance
- This is the first report on the establishment of equine keratinocytes derived from lip epithelium and forms the foundation to study equine keratinocyte biology and the pathogenesis of epidermal diseases.
- Given that wound healing is a major issue in equine dermatology, this research may be crucial for the development of new and improved therapies.
Cite This Article
APA
Dahm AM, de Bruin A, Linat A, von Tscharner C, Wyder M, Suter MM.
(2002).
Cultivation and characterisation of primary and subcultured equine keratinocytes.
Equine Vet J, 34(2), 114-120.
https://doi.org/10.2746/042516402776767187 Publication
Researcher Affiliations
- Institute of Animal Pathology, University of Berne, Switzerland.
MeSH Terms
- Animals
- Cell Differentiation
- Cell Division
- Cells, Cultured
- Collagen
- Horses
- Immunohistochemistry / veterinary
- Keratinocytes / physiology
- Keratinocytes / ultrastructure
- Microscopy, Electron / veterinary
- Skin / cytology
Citations
This article has been cited 3 times.- Schubert AK, Smink JJ, Pumberger M, Putzier M, Sittinger M, Ringe J. Standardisation of basal medium for reproducible culture of human annulus fibrosus and nucleus pulposus cells. J Orthop Surg Res 2018 Aug 22;13(1):209.
- Alkhilaiwi F, Wang L, Zhou D, Raudsepp T, Ghosh S, Paul S, Palechor-Ceron N, Brandt S, Luff J, Liu X, Schlegel R, Yuan H. Long-term expansion of primary equine keratinocytes that maintain the ability to differentiate into stratified epidermis. Stem Cell Res Ther 2018 Jul 4;9(1):181.
- Visser MB, Pollitt CC. Characterization of extracellular matrix macromolecules in primary cultures of equine keratinocytes. BMC Vet Res 2010 Mar 15;6:16.
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