Methods of assessing oxidant injury in renal tubular epithelial cells in vitro

Sharon P. Andreoli, Coleen H. Mallett

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Reactive oxygen molecules mediate renal injury in several diseases including glomerulonephritis, ischemic-reperfusion injury, and toxic nephropathies. Several in vivo animal studies document a role for reactive oxygen molecules in causing tissue injury. In vitro methods to determine the mechanisms and effects of reactive oxygen molecule injury will further aid in understanding the pathogenesis of several renal diseases. We describe methods to study cell injury in an in vitro cell culture system of renal tubular epithelial cells exposed to reactive oxygen molecules generated by hypoxanthine-xanthine oxidase or glucose-glucose oxidase. Methods to determine ATP levels and [3H]adenine release as an early response (15 to 90 min) to oxidant stress are described.51Chromium release representing cell detachment and cell lysis which are late responses (3 to 5 h) are also detailed. Methods of assessing oxidant injury in in vitro systems will allow the precise mechanisms of cell injury to be determined and will allow for the potential development of strategies to prevent or alleviate such injury.

Original languageEnglish (US)
Pages (from-to)199-204
Number of pages6
JournalJournal of tissue culture methods: Tissue Culture Association manual of cell, tissue, and organ culture procedures
Volume13
Issue number3
DOIs
StatePublished - Sep 1991

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Keywords

  • ATP
  • epithelial cells
  • oxidant injury
  • reactive oxygen molecules

ASJC Scopus subject areas

  • Anatomy
  • Pathology and Forensic Medicine

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