title: Xenobiotic- and Serum-Free Culture of Oral Mucosal Epithelial Cells on Contact Lenses.
abstract: OBJECTIVE
  Cultured autologous oral mucosal epithelial cells (OMECs) have proven useful in the treatment of ocular surface disorders. This study is the first to investigate the potential of expanding OMEC in a xenobiotic- and serum-free medium using therapeutic contact lenses (CLs) as a substrate and carrier.
  
  
  METHODS
  Porcine OMEC were seeded on laminin-coated lotrafilcon A therapeutic CLs with the density of 8 × 10(4) cells/lens and cultured in a defined serum and xenobiotic-free medium. Confocal immunofluorescence microscopy was used to analyze the following: (1) cellular morphology by using rhodamine-phalloidin staining of F-actin, (2) phenotype by applying antibodies against the progenitor cell marker p63 and the putative stem cell marker ABCG2 and (3) cell viability by using propidium iodide and Hoechst 33342 dual staining.
  
  
  RESULTS
  Porcine OMEC attached well to the CLs, and cell-to-cell contacts were evident. After three days in culture, the OMEC displayed a confluent monolayer with uniform cobblestone morphology, whereas stratified cultures with 2-3 layers were formed after six days. No significant difference in expression of p63 was observed after three-day culture (79.4 ± 14.8%) compared with six-day culture (60.3 ± 18.9%). ABCG2 expression in the basal cell layer was 6.3 ± 1.0% and 4.8 ± 1.8% after three- and six-day culture, respectively. The basal layer viability of cultured OMECs was 99.3 ± 0.2% and 82.8 ± 1.1% after three and six days culture, respectively.
  
  
  CONCLUSIONS
  The use of therapeutic CLs has potential as a substrate and carrier for OMEC cultured in a xenobiotic- and serum-free culture system.
authors: a Department of Chemistry , Umeå University , Umeå , Sweden .
authors: Benny
authors: B
authors: Björkblom
authors: b Department of Medical Biochemistry , Oslo University Hospital , Oslo , Norway .
authors: Jon R
authors: JR
authors: Eidet
authors: b Department of Medical Biochemistry , Oslo University Hospital , Oslo , Norway .|c Department of Oral Biology, Faculty of Dentistry , University of Oslo , Oslo , Norway .
authors: Tor P
authors: TP
authors: Utheim
authors: d Department of Ophthalmology , Stavanger University Hospital , Stavanger , Norway and.
authors: Harald F
authors: HF
authors: Ulltveit-Moe
authors: d Department of Ophthalmology , Stavanger University Hospital , Stavanger , Norway and.|e The Norwegian Dry Eye Clinic , Oslo , Norway.
authors: Sten
authors: S
authors: Raeder
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002452:Cell Count; D018929:Cell Culture Techniques; D003263:Contact Lenses, Hydrophilic; D016895:Culture Media, Serum-Free; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D020100:Hydrogels; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009061:Mouth Mucosa / Q000166:cytology*; D010641:Phenotype; D012828:Silicones; D034421:Sus scrofa; D015262:Xenobiotics; D005466:Fluorocarbons
chemical_list: D015415:Biomarkers; D016895:Culture Media, Serum-Free; D020100:Hydrogels; D012828:Silicones; D015262:Xenobiotics; C481129:lotrafilcon A; D005466:Fluorocarbons
keywords: Cell culture; contact lens; epithelial cells; oral mucosa