Abstract:
:During the last decade, the unbound glutathione content of cultured adherent cells has become a very important biological marker for many pharmacological and toxicological in vitro studies with regard to the protective role of the tripeptide in its reduced form (GSH). However, the literature does not provide extensive information on the influence of sample preparation on cellular GSH and thiol analyses. Using the fibroblast-like V79 cell line as model, we undertook a comparative study of the efficiency of different procedures reported in the literature with respect to GSH recovery. Depending on the preanalytical step, up to 10-fold discrepancies could be observed in the recovery of intracellular GSH. Different parameters that must be controlled in order to maximize GSH recovery are discussed. The optimal strategy consisted in rapid perchloric acid deproteinization performed directly in the dish, which was extremely valuable for preparing GSH samples from adherent cells, and especially from cells expressing elevated gamma-glutamyl transferase activity.
journal_name
Cell Biol Toxicoljournal_title
Cell biology and toxicologyauthors
Thioudellet C,Oster T,Leroy P,Nicolas A,Wellman Mdoi
10.1007/BF00767495subject
Has Abstractpub_date
1995-04-01 00:00:00pages
103-11issue
2eissn
0742-2091issn
1573-6822journal_volume
11pub_type
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