Abstract
A novel population-balance model was employed to evaluate the suppression of cell death in myeloma NS0 6A1 cells metabolically engineered to over-express the apoptotic suppressor Bcl-2. The model is robust in its ability to simulate cell population dynamics in batch suspension culture and in response to thymidine-induced growth inhibition: 89% of simulated cell concentrations are within two standard deviations of experimental data. Kinetic rate constants in model equations suggest that Bcl-2 over-expression extends culture longevity from 6 days to at least 15 days by suppressing the specific rate of early apoptotic cell formation by more than 6-fold and necrotic cell formation by at least 3-fold, despite nearly a 3-fold decrease in initial cell growth rate and no significant change in the specific rate of late apoptotic cell formation. This computational analysis supports a mechanism in which Bcl-2 is a common mediator of early apoptotic and necrotic events occurring at rates that are dependent on cellular factors accumulating over time. The model has current application to the rational design of cell cultures through metabolic engineering for the industrial production of biopharmaceuticals.
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References
Burlacu A (2003) Regulation of apoptosis by Bcl-2 family proteins. J Cell Mol Med 7:249–257
Chaudhry MA (2006) Bystander effect: biological endpoints and microarray analysis. Mutat Res 597:98–112
Cowger NL, O’Connor KC, Hammond TG, Lacks DJ, Navar GL (1999) Characterization of bimodal cell death of insect cells in a rotating-wall vessel and shaker flask. Biotechnol Bioeng 64:14–26
Figueroa B Jr, Sauerwald TM, Oyler GA, Hardwick JM, Betenbaugh MJ (2003) A comparison of the properties of a Bcl-xL variant to the wild-type anti-apoptosis inhibitor in mammalian cell cultures. Metab Eng 5:230–245
Fussenegger M, Schlatter S, Dätwyler D, Mazur X, Bailey JE (1998) Controlled proliferation by multigene metabolic engineering enhances the productivity of Chinese hamster ovary cells. Nature Biotechnol 16:468–472
Kroemer G, Dallaporta B, Resche-Rigon M (1998) The mitochondrial death/life regulator in apoptosis and necrosis. Annu Rev Physiol 60:619–642
Kroemer G, Reed JC (2000) Mitochondrial control of cell death. Nature Med 6:513–519
O’Connor KC, Cowger NL, De Kee DCR, Schwarz RP (2002) Prolonged shearing of insect cells in a Couette bioreactor. Enzyme Microb Technol 31:600–608
Panetta JC, Evans WE, Cheok MH (2006) Mechanistic mathematical modeling of mercaptopurine effects on cell cycle of human acute lymphoblastic leukaemia cells. Br J Cancer 94:93–100
Simpson NH, Singh RP, Emery AN, Al-Rubeai M (1999) Bcl-2 over-expression reduces growth rate and prolongs G1 phase in continuous chemostat cultures of hybridoma cells. Biotechnol Bioeng 64:174–186
Tey BT, Singh RP, Piredda L, Piacentini M, Al-Rubeai M (2000) Bcl-2 mediated suppression of apoptosis in myeloma NS0 cultures. J Biotechnol 79:147–159
Vives J, Juanola S, Cairó JJ, Gòdia F (2003a) Metabolic engineering of apoptosis in cultured animal cells: implications for the biotechnology industry. Metab Eng 5:124–132
Vives J, Juanola S, Cairó JJ, Prats E, Cornudella L, Gòdia F (2003b) Protective effect of viral homologues of bcl-2 on hybridoma cells under apoptosis-inducing conditions. Biotechnol Prog 19:84–89
Yamabe K, Shimizu S, Kamiike W, Waguri S, Eguchi Y, Hasegawa J-I, Okuno S-I, Yoshioka Y, Ito T, Sawa Y, Uchiyama Y, Tsujimoto Y, Matsuda H (1998) Prevention of hypoxic liver cell necrosis by in vivo human bcl-2 gene transfection. Biochem Biophys Res Commun 243:217–223
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This research was support with a grant from NASA (NAG-9-826).
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O’Connor, K.C., Muhitch, J.W., Lacks, D.J. et al. Modeling suppression of cell death by Bcl-2 over-expression in myeloma NS0 6A1 cells. Biotechnol Lett 28, 1919–1924 (2006). https://doi.org/10.1007/s10529-006-9177-5
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DOI: https://doi.org/10.1007/s10529-006-9177-5