Cargando…

Development and Characterisation of a Human Chronic Skin Wound Cell Line—Towards an Alternative for Animal Experimentation

Background: Chronic skin wounds are a growing financial burden for healthcare providers, causing discomfort/immobility to patients. Whilst animal chronic wound models have been developed to allow for mechanistic studies and to develop/test potential therapies, such systems are not good representatio...

Descripción completa

Detalles Bibliográficos
Autores principales: Caley, Matthew, Wall, Ivan B., Peake, Matthew, Kipling, David, Giles, Peter, Thomas, David W., Stephens, Phil
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5979489/
https://www.ncbi.nlm.nih.gov/pubmed/29584680
http://dx.doi.org/10.3390/ijms19041001
_version_ 1783327709047291904
author Caley, Matthew
Wall, Ivan B.
Peake, Matthew
Kipling, David
Giles, Peter
Thomas, David W.
Stephens, Phil
author_facet Caley, Matthew
Wall, Ivan B.
Peake, Matthew
Kipling, David
Giles, Peter
Thomas, David W.
Stephens, Phil
author_sort Caley, Matthew
collection PubMed
description Background: Chronic skin wounds are a growing financial burden for healthcare providers, causing discomfort/immobility to patients. Whilst animal chronic wound models have been developed to allow for mechanistic studies and to develop/test potential therapies, such systems are not good representations of the human chronic wound state. As an alternative, human chronic wound fibroblasts (CWFs) have permitted an insight into the dysfunctional cellular mechanisms that are associated with these wounds. However, such cells strains have a limited replicative lifespan and therefore a limited reproducibility/usefulness. Objectives: To develop/characterise immortalised cell lines of CWF and patient-matched normal fibroblasts (NFs). Methods and Results: Immortalisation with human telomerase resulted in both CWF and NF proliferating well beyond their replicative senescence end-point (respective cell strains senesced as normal). Gene expression analysis demonstrated that, whilst proliferation-associated genes were up-regulated in the cell lines (as would be expected), the immortalisation process did not significantly affect the disease-specific genotype. Immortalised CWF (as compared to NF) also retained a distinct impairment in their wound repopulation potential (in line with CWF cell strains). Conclusions: These novel CWF cell lines are a credible animal alternative and could be a valuable research tool for understanding both the aetiology of chronic skin wounds and for therapeutic pre-screening.
format Online
Article
Text
id pubmed-5979489
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-59794892018-06-10 Development and Characterisation of a Human Chronic Skin Wound Cell Line—Towards an Alternative for Animal Experimentation Caley, Matthew Wall, Ivan B. Peake, Matthew Kipling, David Giles, Peter Thomas, David W. Stephens, Phil Int J Mol Sci Article Background: Chronic skin wounds are a growing financial burden for healthcare providers, causing discomfort/immobility to patients. Whilst animal chronic wound models have been developed to allow for mechanistic studies and to develop/test potential therapies, such systems are not good representations of the human chronic wound state. As an alternative, human chronic wound fibroblasts (CWFs) have permitted an insight into the dysfunctional cellular mechanisms that are associated with these wounds. However, such cells strains have a limited replicative lifespan and therefore a limited reproducibility/usefulness. Objectives: To develop/characterise immortalised cell lines of CWF and patient-matched normal fibroblasts (NFs). Methods and Results: Immortalisation with human telomerase resulted in both CWF and NF proliferating well beyond their replicative senescence end-point (respective cell strains senesced as normal). Gene expression analysis demonstrated that, whilst proliferation-associated genes were up-regulated in the cell lines (as would be expected), the immortalisation process did not significantly affect the disease-specific genotype. Immortalised CWF (as compared to NF) also retained a distinct impairment in their wound repopulation potential (in line with CWF cell strains). Conclusions: These novel CWF cell lines are a credible animal alternative and could be a valuable research tool for understanding both the aetiology of chronic skin wounds and for therapeutic pre-screening. MDPI 2018-03-27 /pmc/articles/PMC5979489/ /pubmed/29584680 http://dx.doi.org/10.3390/ijms19041001 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Caley, Matthew
Wall, Ivan B.
Peake, Matthew
Kipling, David
Giles, Peter
Thomas, David W.
Stephens, Phil
Development and Characterisation of a Human Chronic Skin Wound Cell Line—Towards an Alternative for Animal Experimentation
title Development and Characterisation of a Human Chronic Skin Wound Cell Line—Towards an Alternative for Animal Experimentation
title_full Development and Characterisation of a Human Chronic Skin Wound Cell Line—Towards an Alternative for Animal Experimentation
title_fullStr Development and Characterisation of a Human Chronic Skin Wound Cell Line—Towards an Alternative for Animal Experimentation
title_full_unstemmed Development and Characterisation of a Human Chronic Skin Wound Cell Line—Towards an Alternative for Animal Experimentation
title_short Development and Characterisation of a Human Chronic Skin Wound Cell Line—Towards an Alternative for Animal Experimentation
title_sort development and characterisation of a human chronic skin wound cell line—towards an alternative for animal experimentation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5979489/
https://www.ncbi.nlm.nih.gov/pubmed/29584680
http://dx.doi.org/10.3390/ijms19041001
work_keys_str_mv AT caleymatthew developmentandcharacterisationofahumanchronicskinwoundcelllinetowardsanalternativeforanimalexperimentation
AT wallivanb developmentandcharacterisationofahumanchronicskinwoundcelllinetowardsanalternativeforanimalexperimentation
AT peakematthew developmentandcharacterisationofahumanchronicskinwoundcelllinetowardsanalternativeforanimalexperimentation
AT kiplingdavid developmentandcharacterisationofahumanchronicskinwoundcelllinetowardsanalternativeforanimalexperimentation
AT gilespeter developmentandcharacterisationofahumanchronicskinwoundcelllinetowardsanalternativeforanimalexperimentation
AT thomasdavidw developmentandcharacterisationofahumanchronicskinwoundcelllinetowardsanalternativeforanimalexperimentation
AT stephensphil developmentandcharacterisationofahumanchronicskinwoundcelllinetowardsanalternativeforanimalexperimentation