Optimizing preincubation conditions for precision-cut rat kidney and liver tissue slices: effect of culture media and antioxidants

Toxicology in Vitro : an International Journal Published in Association with BIBRA
D K ObatomiP H Bach

Abstract

Tissue slices are commonly 'preincubated' before use, but optimal conditions to ensure their maximal viability have not been systematically investigated. The effects of serum-free Dulbecco's minimum Eagle's medium and Ham's nutrient mixture (DMEM/F12) (1:1) culture media with and without phenol red (+/-PR), or RPMI-1640 and six different antioxidants on the viability of precision-cut rat kidney and liver slices (200+/-5mum) were investigated. Slice viability was assessed every 30 minutes over a 2-hour preincubation period and after 24 hours of incubation in a multiwell plate culture system maintained at 37 degrees C. In all cases, preincubation produced a time-dependent significant reduction of ethidium bromide positive nuclei stained in each medium and in both kidney and liver slices. Based on lactate dehydrogenase (LDH) leakage, there are viability differences between the media. In contrast, alkaline phosphatase (ALP) leakage and MTT reduction were less sensitive and did not differentiate between slice viability in each incubation medium. Preincubation of kidney and liver slices in DMEM/F12 medium containing antioxidants, indicated an enhanced viability which was specific for each tissue. Extension of the culture period to 24...Continue Reading

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Citations

Aug 29, 2001·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·D K ObatomiP H Bach
Nov 25, 2000·Toxicology·C Lerche-Langrand, H J Toutain
Jan 23, 2003·Toxicology in Vitro : an International Journal Published in Association with BIBRA·I A M de Graaf, H J Koster
Oct 31, 2000·Xenobiotica; the Fate of Foreign Compounds in Biological Systems·A K Sohlenius-SternbeckA Orzechowski
Apr 19, 2007·Clinical and Experimental Pharmacology & Physiology·Yu GuoChun Zhang
Jan 29, 2011·Biotechnology and Bioengineering·Paul M van MidwoudElisabeth Verpoorte
Sep 1, 2009·Biotechnology and Bioengineering·Paul M van MidwoudElisabeth Verpoorte
Jan 27, 2015·Nitric Oxide : Biology and Chemistry·Pan HuangHongfang Jin
Mar 21, 2017·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Fandi KongXiaoyan Chen
Dec 7, 2010·Analytical Chemistry·Paul M van MidwoudElisabeth Verpoorte

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