title: The protection of hybridoma cells from apoptosis by caspase inhibition allows culture recovery when exposed to non-inducing conditions.
abstract: Programmed cell death (PCD) or apoptosis process in a hybridoma cell line induced by the deprivation of one of the main nutrients, glutamine, has been studied. The use of caspase inhibitors has enabled maintenance of cell viability during a significant period of time, when glutamine depletion was maintained in the culture. Two caspase inhibitors partially suppressed the progress of PCD under glutamine deprivation: Ac-DEVD-cho and z-VAD-fmk. Indeed, as a consequence of this protection, the number of viable cells decreased by 10% (for z-VAD-fmk) and by 80% (for Ac-DEVD-cmk) after 36 h of culture, while it decreased by 90% for a control culture in the absence of protective compounds. However, when the culture was exposed to non-apoptotic conditions after this period of time under apoptosis protection conditions, a normal growth pattern was not recovered. Interestingly, the simultaneous use of both inhibitors made the recovery of the cell culture possible even after a period of 36 h under glutamine depletion, indicating that the inhibition of the effector caspases occurs upstream of the point in which hybridoma cells enter into the commitment step of the death programme.
authors: Dept. d'Enginyeria Química, Universitate Autonoma de Barcelona, Escola Tècnica Superior d'Enginyers, ETSE, 08193 Bellaterra, Barcelona, Spain.
authors: Albert
authors: A
authors: Tintó
authors: Carme
authors: C
authors: Gabernet
authors: Joaquim
authors: J
authors: Vives
authors: Eva
authors: E
authors: Prats
authors: Jordi J
authors: JJ
authors: Cairó
authors: Lluís
authors: L
authors: Cornudella
authors: Francesc
authors: F
authors: Gòdia
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D061945:Caspase Inhibitors; D018929:Cell Culture Techniques; D002455:Cell Division; D002470:Cell Survival; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D005973:Glutamine / Q000378:metabolism; D006825:Hybridomas / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D051379:Mice; D009842:Oligopeptides / Q000494:pharmacology
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D009842:Oligopeptides; C099158:acetyl-aspartyl-glutamyl-valyl-aspartal; C459419:benzyloxycarbonyltyrosyl-alanyl-valyl-aspartyl chloromethyl ketone; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D005973:Glutamine