title: LLC-PK(1) cells maintained in a new perfusion cell culture system exhibit an improved oxidative metabolism.
abstract: 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.
authors.affiliation: Dept. of Cell and Developmental Biology, University of Pennsylvania, USA.
authors.forename: Edward
authors.initials: E
authors.lastname: Felder
authors.forename: Paul
authors.initials: P
authors.lastname: Jennings
authors.forename: Thomas
authors.initials: T
authors.lastname: Seppi
authors.forename: Walter
authors.initials: W
authors.lastname: Pfaller
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002457:Cell Extracts; D048429:Cell Size; D003470:Culture Media / Q000737:chemistry; D005947:Glucose / Q000378:metabolism; D006019:Glycolysis; D018374:LLC-PK1 Cells; D019344:Lactic Acid / Q000378:metabolism; D008854:Microscopy, Electron; D010100:Oxygen / Q000032:analysis / Q000378:metabolism*; D010477:Perfusion / Q000295:instrumentation* / Q000379:methods*; D013552:Swine
chemical_list: D002457:Cell Extracts; D003470:Culture Media; D019344:Lactic Acid; D005947:Glucose; D010100:Oxygen