Insulin and transferrin restore important cellular functions of human fetal kidney in serum-free organ culture.
A model previously developed in our laboratory to culture human fetal kidneys in serum-free chemically defined medium was used to evaluate the direct influence of potential regulators on nephrogenesis. The aim of the present work was to verify the effects of insulin and transferrin, two hormones considered as essential in other serum-free culture systems. Explants of renal cortex from human fetuses (15-21 weeks) were cultured for 2 and 5 days in serum-free Leibovitz's L-15 medium (37 degrees C, 95% air - 5% CO2). The addition of transferrin (5 micrograms/mL) had no effect, but insulin (30, 60, and 125 mU/mL) increased DNA and protein syntheses in a dose-dependent manner. The influence of insulin (125 mU/mL) was potentiated by the addition of transferrin and the combination of the two stimulated DNA synthesis by threefold on day 2 when compared with controls and by sixfold on day 5 of culture. After 5 days, synthesis was restored to values observed at day 0. Transferrin did not modify the insulin effect on protein synthesis, since the latter was already maximally stimulated as early as day 2 of culture and at levels well above that of uncultured explants (day 0). The activities of four hydrolases considered as markers of brush border differentiation were not importantly changed by any of the hormones, supplemented alone or in combination. The results indicate that proliferation rather than differentiation is the parameter mostly influenced by these two hormones. The combination of insulin plus transferrin restores cellular functions of human fetal kidney explants cultured in serum-free medium.(ABSTRACT TRUNCATED AT 250 WORDS)
Brière
N
N
Département d'anatomie et de biologie cellulaire, Faculté de médecine, Université de Sherbrooke, Québec, Canada.
Ferrari
J
J
Chailler
P
P
D000469:Alkaline Phosphatase / Q000378:metabolism; D000581:Amidohydrolases / Q000378:metabolism; D003470:Culture Media; D004247:DNA / Q000096:biosynthesis; D005333:Fetus; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000196:embryology* / Q000254:growth & development; D009924:Organ Culture Techniques; D014176:Protein Biosynthesis; D014168:Transferrin / Q000494:pharmacology*; D000520:alpha-Glucosidases / Q000378:metabolism; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D003470:Culture Media; D007328:Insulin; D014168:Transferrin; D004247:DNA; D005723:gamma-Glutamyltransferase; D000469:Alkaline Phosphatase; D000520:alpha-Glucosidases; D000581:Amidohydrolases