Year-end Sale % OFF 
Abstract
BackgroundSkin diseases are a major health problem. Some of the most severe conditions involve genetic disorders, including cancer. Several of these human diseases have been modelled in genetically modified mice, thus becoming a highly valuable preclinical tool for the treatment of these pathologies. However, development of three-dimensional models of skin using keratinocytes from normal and/or genetically modified mice has been hindered by the difficulty to subculture murine epidermal keratinocytes.MethodsWe have generated a murine epidermal cell line by serially passaging keratinocytes isolated from the back skin of adult mice. We have termed this cell line COCA. Cell culture is done in fully defined media and does not require feeder cells or any other coating methods.ResultsCOCA retained its capacity to differentiate and stratify in response to increased calcium concentration in the cell culture medium for more than 75 passages. These cells, including late passage, can form epidermis-like structures in three-dimensional in vitro models with a well-preserved pattern of proliferation and differentiation. Furthermore, these cells form epidermis in grafting assays in vivo, and do not develop tumorigenic ability.ConclusionsWe propose that COCA constitutes a good experimental system for in vitro and in vivo skin modelling. Also, cell lines from genetically modified mice of interest in skin biology could be established using the method we have developed. COCA keratinocytes would be a suitable control, within a similar background, when studying the biological implications of these alterations.
Highlights
Skin diseases are a major health problem
Establishment of a long-term culture of murine epidermal keratinocytes Keratinocytes were isolated from the back skin of 2months old C57BL/DBA mice and maintained in culture in low calcium (0.07 mM) fully defined media (CnT-07)
Cells were repeatedly passaged to establish a cell line. We termed this murine epidermal keratinocyte derived cell line COCA, and we have expanded it in culture for more than 80 passages
Summary
Some of the most severe conditions involve genetic disorders, including cancer. Several of these human diseases have been modelled in genetically modified mice, becoming a highly valuable preclinical tool for the treatment of these pathologies. Development of three-dimensional models of skin using keratinocytes from normal and/or genetically modified mice has been hindered by the difficulty to subculture murine epidermal keratinocytes. Skin diseases affect patients of all ages worldwide By some estimates, they affect 50 percent or more of the population at any one time and comprise over 2,000 medical conditions which can range from only mild skin problems to pathologies that are serious or even fatal. The aetiology of skin diseases is varied and some of the most severe are autoimmune and genetic disorders, including cancer.
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
