Modulation of glycolysis induction in primary cultures of rabbit kidney proximal tubule cells: the role of shaking, glucose and insulin.
We have assessed the impact of increasing oxygen availability on cellular phenotype expression of rabbit proximal tubule cells in primary culture developed with variable glucose and/or insulin contents. To mitigate hypoxia at the cell/medium interface, cells were shaken for the whole culture duration and their expressed phenotype was compared with those expressed by static cultures. O2 and CO2 tensions were kept constant in the incubator atmosphere. Glycolysis and gluconeogenesis pathways, detoxication system, and mitochondrial, apical and basolateral membrane marker enzyme activities were assessed. This study showed that the induction of glycolysis which appear in primary cultures of proximal tubule cells may be partially prevented by continuously shaking the cultures. This effect was more marked in the presence of glucose, suggesting better substrate oxidation in shaken cultures.
Monteil
C
C
INSERM U-295, Universite de Rouen, Saint Etienne du Rouvray, France.
Leclere
C
C
Dantzer
F
F
Elkaz
V
V
Fillastre
J P
JP
Morin
J P
JP
D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000641:Ammonia / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D003470:Culture Media; D005943:Gluconeogenesis; D005947:Glucose / Q000378:metabolism*; D005978:Glutathione / Q000096:biosynthesis; D006019:Glycolysis; D007328:Insulin / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D007773:Lactates / Q000378:metabolism; D019344:Lactic Acid; D010101:Oxygen Consumption; D011817:Rabbits
D003470:Culture Media; D007328:Insulin; D007773:Lactates; D019344:Lactic Acid; D000641:Ammonia; D000255:Adenosine Triphosphate; D005978:Glutathione; D005947:Glucose