Alteration of culture regime modifies antioxidant defenses independent of intracellular reactive oxygen levels and resistance to severe oxidative stress within confluent Caco-2 "intestinal cells".
Reactive oxygen species are implicated in the development of gastrointestinal pathologies. Caco-2 monolayers are routinely used to study intestinal oxidative stress and its potential amelioration by pharmacological agents or dietary micronutrients. Little is known of the plasticity of Caco-2 antioxidant defenses with changes in culture conditions. We examined whether the frequency of culture media renewal alters the antioxidant-prooxidant status and integrity of Caco-2 monolayers. In comparison to monolayers subject to daily media renewal, increasing periods between media exchange resulted in varying degrees of suppression of catalase, glutathione reductase, and glutathione-S-transferase activity. No significant changes to superoxide dismutase activity, total glutathione, or intracellular ROS profiles were observed. Alkaline phosphatase activity, as a marker of differentiation, and mean monolayer cell population size were also unaffected. We suggest that Caco-2 antioxidant enzyme activities are differentially sensitive to changes in culture conditions. Studies employing this cell line for antioxidant-oxidative stress interactions will need to evaluate responses with respect to culture regime.
Bestwick
C S
CS
Antioxidant and DNA Damage Group, Cellular Integrity Programme, Rowett Research Institute, Aberdeen, UK.
Milne
L
L
D000469:Alkaline Phosphatase / Q000032:analysis; D000975:Antioxidants / Q000494:pharmacology*; D018938:Caco-2 Cells / Q000737:chemistry / Q000187:drug effects* / Q000276:immunology* / Q000378:metabolism; D002374:Catalase / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media / Q000737:chemistry* / Q000378:metabolism; D005978:Glutathione / Q000032:analysis; D005980:Glutathione Reductase / Q000032:analysis; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000276:immunology*; D018384:Oxidative Stress / Q000187:drug effects* / Q000276:immunology*; D017382:Reactive Oxygen Species / Q000276:immunology* / Q000378:metabolism; D013482:Superoxide Dismutase / Q000032:analysis; D013997:Time Factors
D000975:Antioxidants; D003470:Culture Media; D017382:Reactive Oxygen Species; D002374:Catalase; D013482:Superoxide Dismutase; D005980:Glutathione Reductase; D000469:Alkaline Phosphatase; D005978:Glutathione