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"

Charles Bestwick, Lesley Milne

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

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 chan ses 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.

Original languageEnglish
Pages (from-to)417-423
Number of pages7
JournalDigestive Diseases and Sciences
Volume46
Issue number2
DOIs
Publication statusPublished - Feb 2001

Keywords

  • cell culture
  • intestine
  • antioxidant enzymes
  • reactive oxygen
  • oxidative stress
  • Caco-2 cells
  • free-radicals
  • line CACO-2
  • human-colon
  • gene-expression
  • differentiation
  • superoxide
  • enzymes
  • disease

Cite this

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title = "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{"}",
abstract = "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 chan ses 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.",
keywords = "cell culture, intestine, antioxidant enzymes, reactive oxygen, oxidative stress, Caco-2 cells, free-radicals, line CACO-2, human-colon, gene-expression, differentiation, superoxide, enzymes, disease",
author = "Charles Bestwick and Lesley Milne",
year = "2001",
month = "2",
doi = "10.1023/A:1005621403626",
language = "English",
volume = "46",
pages = "417--423",
journal = "Digestive Diseases and Sciences",
issn = "0163-2116",
publisher = "Springer New York",
number = "2",

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TY - JOUR

T1 - 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"

AU - Bestwick, Charles

AU - Milne, Lesley

PY - 2001/2

Y1 - 2001/2

N2 - 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 chan ses 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.

AB - 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 chan ses 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.

KW - cell culture

KW - intestine

KW - antioxidant enzymes

KW - reactive oxygen

KW - oxidative stress

KW - Caco-2 cells

KW - free-radicals

KW - line CACO-2

KW - human-colon

KW - gene-expression

KW - differentiation

KW - superoxide

KW - enzymes

KW - disease

U2 - 10.1023/A:1005621403626

DO - 10.1023/A:1005621403626

M3 - Article

VL - 46

SP - 417

EP - 423

JO - Digestive Diseases and Sciences

JF - Digestive Diseases and Sciences

SN - 0163-2116

IS - 2

ER -