Establishment and characterization of a canine keratinocyte organoid culture system.

Wiener, Dominique Judith; Basak, Onur; Asra, Priyanca; Boonekamp, Kim E; Kretzschmar, Kai; Papaspyropoulos, Angelos; Clevers, Hans (2018). Establishment and characterization of a canine keratinocyte organoid culture system. Veterinary dermatology, 29(5), 375-e126. Blackwell Science 10.1111/vde.12541

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BACKGROUND Perturbations of epidermal and follicular homeostasis have been attributed to a variety of skin diseases affecting dogs. The availability of an in vitro system to investigate these diseases is important to understand underlying pathomechanisms. OBJECTIVES To establish an accurate and reliable in vitro 3D system of canine keratinocyte organoids to lay the basis for studying functional defects in interfollicular epidermis (IFE) and hair follicle (HF) morphogenesis, reconstitution and differentiation that lead to alopecic and epidermal diseases. ANIMALS Skin biopsies were obtained from freshly euthanized dogs of different breeds with no skin abnormalities. METHODS Cells derived from microdissected IFE and HFs were seeded in Matrigel and keratinocyte organoids were grown and characterized using immunohistochemistry, RT-qPCR and RNA sequencing. RESULTS Both organoid lines develop into a basal IFE-like cell type. Gene and protein expression analysis revealed high mRNA and protein levels of keratins 5 and 14, IFE differentiation markers and intercellular molecules. Key markers of HF stem cells were lacking. Withdrawal of growth factors resulted in upregulation of markers such as KRT16, Involucrin, KRT17 and SOX9, showing the potential of the organoids to develop towards more differentiated tissue. CONCLUSION AND CLINICAL IMPORTANCE Our 3D in vitro culture system provides the basis to explore epidermal function, to investigate the culture conditions necessary for the development of organoids with a HF signature and to address cutaneous disorders in dogs. However, for induction of HF signatures or hair growth, addition of different growth factors or co-culture with dermal papilla will be required.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > DermFocus
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Animal Pathology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)

UniBE Contributor:

Wiener, Dominique Judith

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 590 Animals (Zoology)
600 Technology > 630 Agriculture

ISSN:

0959-4493

Publisher:

Blackwell Science

Language:

English

Submitter:

Achim Braun Parham

Date Deposited:

03 Jun 2019 11:20

Last Modified:

24 Oct 2019 09:32

Publisher DOI:

10.1111/vde.12541

PubMed ID:

29963730

BORIS DOI:

10.7892/boris.127541

URI:

https://boris.unibe.ch/id/eprint/127541

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