Metabolic viability and pharmaco-toxicological reactivity of cryopreserved human precision-cut renal cortical slices.
We have tested the suitability of cryopreserved human precision-cut renal cortical slices for metabolic and pharmaco-toxicological studies. The viability of these slices and their pharmaco-toxicological reactivity were assessed using intracellular ATP and protein contents, lactate dehydrogenase (LDH) leakage, lactate and glutamine metabolism and the ammoniagenic effect of valproate. Despite a decrease in ATP and protein contents when compared with those of fresh slices, cryopreserved slices did not show any LDH leakage and retained the capacity to metabolize glutamine and lactate. Glutamine removal and ammonia, lactate and alanine production were similar in fresh and cryopreserved slices; by contrast, cryopreserved slices accumulated more glutamate as a result of decreased flux through glutamate dehydrogenase which catalyses an oxygen-dependent reaction. Valproate markedly and similarly stimulated glutamine metabolism in fresh and cryopreserved slices. Cryopreservation did not alter lactate removal but inhibited lactate gluconeogenesis. In conclusion, these results demonstrate that, although their mitochondrial oxidative metabolism seems to be diminished, cryopreserved human precision-cut renal cortical slices remain metabolically viable and retain the capacity to respond to the ammoniagenic effect of valproate. Thus, this experimental model may be helpful to optimize the use of human renal tissue for metabolic and pharmaco-toxicological studies.
Vittorelli
Anne
A
Laboratoire de Physiopathologie Métabolique et Rénale, Institut National de la Santé et de la Recherche Médicale, U 499, Faculté de Médecine R.T.H. Laennec, Lyon, France.
Gauthier
Catherine
C
Michoudet
Christian
C
Baverel
Gabriel
G
D000255:Adenosine Triphosphate / Q000032:analysis; D002250:Carbon Radioisotopes; D002470:Cell Survival; D015925:Cryopreservation; D005973:Glutamine / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007672:Kidney Cortex / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D011506:Proteins / Q000032:analysis; D013997:Time Factors; D014635:Valproic Acid / Q000494:pharmacology
D002250:Carbon Radioisotopes; D011506:Proteins; D005973:Glutamine; D014635:Valproic Acid; D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase