LLC-PK(1) cells maintained in a new perfusion cell culture system exhibit an improved oxidative metabolism

Edward Felder, Paul Jennings, Thomas Seppi, Walter Pfaller

Research output: Contribution to JournalArticleAcademicpeer-review


Cultured renal proximal tubule cells dedifferentiate from an oxidative metabolism to high rates of glycolysis over time. There are many reasons why cells in culture dedifferentiate, not least being a lack of homogenous nutrient supply and poor oxygenation. To this end we have developed a new cell culture device (EpiFlow), which combines continuous perfusion of medium with continuous oxygenation of cells grown on microporous supports. LLC-PK(1) cells cultured under EpiFlow conditions were compared with the same cells grown under conventional static conditions. EpiFlow maintained cells exhibited an improved oxidative metabolism as evidenced by 1) a decreased activity of glycolytic enzymes, 2) an increase in the activity of mitochondrial phosphate-dependent-glutaminase, 3) an increase in cellular ATP content, and 4) an improved morphology (increased cell height, mitochondrial density and an increased number and height of microvilli). In addition, LLC-PK(1) cells maintained under perfusion conditions exhibited an increased sensitivity to the respiratory chain blocker antimycin A as assayed by mitochondrial membrane potential (JC-1). We conclude that LLC-PK(1) cells maintained under EpiFlow conditions develop an improved oxidative metabolism that is more comparable to the in vivo situation.

Original languageEnglish
Pages (from-to)153-62
Number of pages10
JournalCellular Physiology and Biochemistry
Issue number2-3
Publication statusPublished - 2002


  • Animals
  • Cell Culture Techniques
  • Cell Extracts
  • Cell Size
  • Culture Media
  • Glucose
  • Glycolysis
  • LLC-PK1 Cells
  • Lactic Acid
  • Microscopy, Electron
  • Oxygen
  • Perfusion
  • Swine
  • Journal Article


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