Development and characterization of an equine skin-equivalent model.
Abstract: There is increasing interest in the biological and pathological study of equine skin owing to the high prevalence of cutaneous diseases in horses. However, knowledge of equine skin cell biology and cultures is limited by the low number of in vitro studies in the literature. Objective: The objective of the study was to develop and characterize an in vitro equine skin equivalent. Methods: Cultures of pure equine keratinocytes and dermal fibroblasts were obtained by enzymatic digestion of skin biopsies. Fibroblasts were embedded into type I collagen matrices to obtain dermal scaffolds, the surface of which was seeded with keratinocytes. The three-dimensional cultures were exposed to the air-liquid interface to enable epidermal stratification. Results: After 14 days in air-exposed conditions, histological analysis showed that keratinocytes underwent differentiation into a multilayered epidermis. Immunohistochemical studies revealed the expression of epidermal cytokeratin in keratinocytes, whereas vimentin was expressed in dermal fibroblasts, as expected in equine skin. Immunostaining of Ki67 showed proliferative keratinocytes in the stratum basale. A continuous basement membrane at the dermo-epidermal junction was also detected immunohistochemically through the expression of its major components (type IV collagen and laminin 5). Ultrastructural analysis by electron microscopy showed desmosomes located among keratinocytes in all layers and hemidesmosomes among the basal keratinocytes and lamina densa. Conclusions: This study reports, for the first time, the development of an in vitro equine skin-equivalent model that resembles equine skin morphologically, immunohistochemically and ultrastructurally.
© 2014 ESVD and ACVD.
Publication Date: 2014-07-18 PubMed ID: 25041278DOI: 10.1111/vde.12134Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research study focused on developing and characterizing an in vitro model of equine skin due to the increasing interest in studying equine skin diseases. It showcases the development of a three-dimensional model with characteristics closely resembling real horse skin.
Objective and Methodology
- The researchers aimed to create an in vitro (laboratory-made) model of equine skin due to the many skin diseases that affect horses. This new model could allow for better understanding and studying of equine skin cell biology.
- Equine keratinocytes and dermal fibroblasts, two types of skin cells, were obtained through enzymatic digestion of skin biopsies. Dermal fibroblasts were embedded into type I collagen matrices to create dermal scaffolds.
- Keratinocytes were seeded on these scaffolds, and this setup was then exposed to an air-liquid interface, a process which guides the growth of the keratinocytes into a multilayered epidermis, similar to the outer layer of skin.
Findings
- The findings revealed that, after 14 days of air-exposed conditions, the keratinocytes evolved into a multilayered epidermis, imitating the structure of the skin.
- The immunohistochemical studies conducted as part of the research showed distinctive markers for both types of cells, with keratinocytes expressing cytokeratin and fibroblasts expressing vimentin. These cell expressions are found in actual equine skin too.
- Additionally, a basement membrane forming the boundary between the epidermis and the dermis was also detected. This membrane expressed major components typical of real skin: type IV collagen and laminin 5.
- Finally, an ultrastructural analysis revealed the presence of desmosomes, structures that provide adhesion between cells, across all layers and hemidesmosomes, which attach epithelial cells to the basement membrane, between the basal keratinocytes and lamina densa.
Conclusion
- This research, for the first time, reported the successful development of an in vitro equine skin-equivalent model that closely mimics actual equine skin, not just morphologically but also immunohistochemically and ultrastructurally.
- This model can have numerous applications in studying the pathological conditions of equine skin and the biology of skin cells.
Cite This Article
APA
Cerrato S, Ramió-Lluch L, Brazís P, Rabanal RM, Fondevila D, Puigdemont A.
(2014).
Development and characterization of an equine skin-equivalent model.
Vet Dermatol, 25(5), 475-e77.
https://doi.org/10.1111/vde.12134 Publication
Researcher Affiliations
- UNIVET, Edifici Astrolabio, Avinguda Cerdanyola 92, 08172, Sant Cugat del Vallès, Barcelona, Spain.
MeSH Terms
- Animals
- Cell Culture Techniques / veterinary
- Collagen
- Fibroblasts / physiology
- Horses / anatomy & histology
- Keratinocytes / physiology
- Skin / anatomy & histology
- Skin / ultrastructure
Citations
This article has been cited 6 times.- Zamith Cunha R, Zannoni A, Salamanca G, De Silva M, Rinnovati R, Gramenzi A, Forni M, Chiocchetti R. Expression of cannabinoid (CB1 and CB2) and cannabinoid-related receptors (TRPV1, GPR55, and PPARα) in the synovial membrane of the horse metacarpophalangeal joint.. Front Vet Sci 2023;10:1045030.
- Ramsauer AS, Wachoski-Dark GL, Fraefel C, Ackermann M, Brandt S, Grest P, Knight CG, Favrot C, Tobler K. Establishment of a Three-Dimensional In Vitro Model of Equine Papillomavirus Type 2 Infection.. Viruses 2021 Jul 19;13(7).
- Souci L, Denesvre C. 3D skin models in domestic animals.. Vet Res 2021 Feb 15;52(1):21.
- Zhao R, Yihan W, Zhao Y, Li B, Han H, Mongke T, Bao T, Wang W, Dugarjaviin M, Bai D. Hair follicle regional specificity in different parts of bay Mongolian horse by histology and transcriptional profiling.. BMC Genomics 2020 Sep 22;21(1):651.
- Work TM, Dagenais J, Weatherby TM, Balazs GH, Ackermann M. In Vitro Replication of Chelonid Herpesvirus 5 in Organotypic Skin Cultures from Hawaiian Green Turtles (Chelonia mydas).. J Virol 2017 Sep 1;91(17).
- Linardi RL, Megee SO, Mainardi SR, Senoo M, Galantino-Homer HL. Expression and localization of epithelial stem cell and differentiation markers in equine skin, eye and hoof.. Vet Dermatol 2015 Aug;26(4):213-e47.
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