title: Differential responses of choroidal melanocytes and uveal melanoma cells to low oxygen conditions.
abstract: Tissue culture is traditionally performed at atmospheric oxygen concentration (21%), which induces hyperoxic stress, as endogenous physiologic oxygen tension found in tissues varies between 2% and 9%. This discrepancy may lead to misinterpretation of results and may explain why effects observed in vitro cannot always be reproduced in vivo and vice versa. Only a few studies have been conducted in low physiologic oxygen conditions to understand the development and differentiation of cells from the eye.
  
  
  
  The aim of this study was to investigate the growth and gene expression profile of melanocytes from the choroid permanently exposed to 21% (hyperoxic) or 3% (physiologic) oxygen with proliferation assays and DNA microarray. The cellular behavior of the melanocytes was then compared to that of cancer cells.
  
  
  
  The gross morphology and melanin content of choroidal melanocytes changed slightly when they were exposed to 3% O2, and the doubling time was statistically significantly faster. There was an increase in the percentage of choroidal melanocytes in the active phases of the cell cycle as observed by using the proliferation marker Ki67. The caveolin-1 senescence marker was not increased in choroidal melanocytes or uveal melanoma cells grown in hyperoxia. In comparison, the morphology of the uveal melanoma cells was similar between the two oxygen levels, and the doubling time was slower at 3% O2. Surprisingly, gene expression profiling of the choroidal melanocytes did not reveal a large list of transcripts considerably dysregulated between the two oxygen concentrations; only the lactate transporter monocarboxylate transporter (MCT4) was statistically significantly upregulated at 3% O2.
  
  
  
  This study showed that the oxygen concentration must be tightly controlled in experimental settings, because it influences the subsequent cellular behavior of human choroidal melanocytes.
authors.affiliation: Axe Médecine régénératrice and Centre Universitaire d'Ophtalmologie (CUO)-Recherche, Centre de recherche du CHU de Québec, Quebec City, QC, Canada; Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Quebec City, QC, Canada.
authors.forename: Cindy
authors.initials: C
authors.lastname: Weidmann
authors.affiliation: Axe Médecine régénératrice and Centre Universitaire d'Ophtalmologie (CUO)-Recherche, Centre de recherche du CHU de Québec, Quebec City, QC, Canada; Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Quebec City, QC, Canada.
authors.forename: Jade
authors.initials: J
authors.lastname: Pomerleau
authors.affiliation: Axe Médecine régénératrice and Centre Universitaire d'Ophtalmologie (CUO)-Recherche, Centre de recherche du CHU de Québec, Quebec City, QC, Canada; Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX, Quebec City, QC, Canada.
authors.forename: Laurence
authors.initials: L
authors.lastname: Trudel-Vandal
authors.affiliation: Axe Médecine régénératrice and Centre Universitaire d'Ophtalmologie (CUO)-Recherche, Centre de recherche du CHU de Québec, Quebec City, QC, Canada; Département d'ophtalmologie, Faculté de médecine, Université Laval, Quebec City, QC, Canada.
authors.forename: Solange
authors.initials: S
authors.lastname: Landreville
mesh_terms: D000369:Aged, 80 and over; D002452:Cell Count; D053145:Cell Nucleus Size / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002829:Choroid / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D019344:Lactic Acid / Q000378:metabolism; D008543:Melanins / Q000096:biosynthesis; D008544:Melanocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D008545:Melanoma / Q000473:pathology*; D008875:Middle Aged; D027501:Monocarboxylic Acid Transporters / Q000235:genetics / Q000378:metabolism; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014604:Uveal Neoplasms / Q000473:pathology*; D000098943:Uveal Melanoma
chemical_list: D008543:Melanins; D027501:Monocarboxylic Acid Transporters; D009124:Muscle Proteins; D012333:RNA, Messenger; C406962:SLC16A4 protein, human; D019344:Lactic Acid; D010100:Oxygen