title: Establishment of a murine epidermal cell line suitable for in vitro and in vivo skin modelling.
abstract: BACKGROUND
  Skin 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.
  
  
  METHODS
  We 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.
  
  
  RESULTS
  COCA 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.
  
  
  CONCLUSIONS
  We 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.
authors.affiliation: Molecular Oncology Unit, Epithelial Biomedicine Division, Basic Research Department, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT), Madrid 28040, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Segrelles
authors.forename: Almudena
authors.initials: A
authors.lastname: Holguín
authors.forename: Pilar
authors.initials: P
authors.lastname: Hernández
authors.forename: José M
authors.initials: JM
authors.lastname: Ariza
authors.forename: Jesús M
authors.initials: JM
authors.lastname: Paramio
authors.forename: Corina
authors.initials: C
authors.lastname: Lorz
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line / Q000187:drug effects / Q000637:transplantation; D003433:Crosses, Genetic; D003470:Culture Media; D000078404:Epidermal Cells; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D007621:Karyotyping; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008819:Mice, Nude; D011123:Polyploidy; D014178:Translocation, Genetic
chemical_list: D003470:Culture Media; D002118:Calcium