Protective effect of viral homologues of bcl-2 on hybridoma cells under apoptosis-inducing conditions.
Targets for metabolic engineering have been identified in a hybridoma cell line to make it more robust in culture toward potential limitations inducing apoptosis. The cells were genetically modified with plasmids harboring endogenous bcl-2 gene and also with viral Bcl-2 homologues, particularly ksbcl-2 and bhrf-1 genes. When cells were exposed to apoptosis-inducing conditions (i.e., glutamine-free medium), the control cells exhibited a decrease in viable cell number within the first 12 h, whereas, for the bcl-2 and ksbcl-2 transfected cell cultures, the viable cell number did not exhibit any clear decrease until after 60 h. Furthermore, hybridoma cells expressing the viral homologue bhrf-1 were even more resistant to cell death, showing a decrease in viability of only 50% at 72 h of culture in glutamine-deprived medium, substantially lower than the 90% viability decrease observed for the control culture. In addition, and most relevant for further bioprocess applications, the cells genetically modified could be brought back to growth conditions even after being exposed to glutamine-deprived conditions during a significant time window, up to 72 h.
Vives
Joaquim
J
Departament d'Enginyeria Química, Escola Tècnica Superior d'Enginyeria (ETSE), Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain.
Juanola
Sandra
S
Cairó
Jordi J
JJ
Prats
Eva
E
Cornudella
Lluís
L
Gòdia
Francesc
F
D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology*; D005818:Genetic Engineering / Q000379:methods*; D005973:Glutamine / Q000494:pharmacology; D006825:Hybridomas / Q000166:cytology / Q000502:physiology*; D051379:Mice; D020011:Protective Agents / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology
D020011:Protective Agents; D019253:Proto-Oncogene Proteins c-bcl-2; D011994:Recombinant Proteins; D014764:Viral Proteins; D005973:Glutamine