Rabbit kidney proximal tubule cells in primary culture: Evaluation of the impact of expressed phenotype on cellular toxic response.
A primary culture of rabbit kidney proximal tubule cells has been developed from highly purified and calibrated fragments of proximal tubules. Cells are grown without serum in various media. Minimum medium (MM) was composed of glucose-free Dulbecco's modified Eagle's medium plus Ham's F12 (1:1) fortified with sodium selenite plus hydrocortisone plus transferrin. The five types of culture medium used for this study were MM plus 15 mm-glucose with or without insulin (media Ins(+) G1 15 mm; Ins(-) G1 15 mm), MM plus 2.5 mm-glucose, and MM with or without insulin (Ins(+) G1(-); Ins(-) G1(-)). Cellular phenotype is characterized by the assessment of (a) the specific activities of eight enzymatic markers, and (b) cellular functions such as DNA and protein synthesis, and methylglucose transport. This study emphasizes the importance of the effect of the composition of the culture medium on the quality of the phenotype expressed by proximal tubule cells in primary culture. In addition, a study involving the well-known nephrotoxic agent gentamicin has demonstrated the impact of the expressed phenotype on the onset and expression of the specific toxicity pattern: the best gentamicin-toxicity pattern is seen in a medium deprived of insulin and glucose.
Morin
J P
JP
INSERM U-295, Université de Rouen, BP 97, 76800 Saint Etienne du Rouvray, France.
Monteil
C
C
Glomot
R
R
Fillastre
J P
JP