The protection of hybridoma cells from apoptosis by caspase inhibition allows culture recovery when exposed to non-inducing conditions.
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.
Tintó
Albert
A
Dept. d'Enginyeria Química, Universitate Autonoma de Barcelona, Escola Tècnica Superior d'Enginyers, ETSE, 08193 Bellaterra, Barcelona, Spain.
Gabernet
Carme
C
Vives
Joaquim
J
Prats
Eva
E
Cairó
Jordi J
JJ
Cornudella
Lluís
L
Gòdia
Francesc
F
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
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