Abstract
COS, myeloma and HeLa cells, which are commonly used for protein production by cell culture, were transfected with human bcl-2 gene encoded on the shuttle vector BCMGS. Expression of human bcl-2 improved survival of cells remarkably, mildly, or negligibly for COS, myeloma, and HeLa, respectively. Four clones were obtained from the human bcl-2 expressing cell population of COS cells. They expressed human bcl-2 almost at the same level. The viable cell numbers were 6, 2.5, 2.5, and 0.8 times as many for the clones #8, #5, #6, and #7, respectively, as for the control COS cells, when they were cultured at low (0.2%) serum concentration for 9 days. The bcl-2 overexpressing COS cells showed morphology different from that of the control COS cells in serum limited condition. When transfected with mouse lambda protein gene carried by an SV40-derived vector, clone #8 of the bcl-2 transfected COS cells continued the transient expression of lambda protein longer than the control COS cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ausbel FM, Brent R, Kingstone RE, Moors DD, Seidman JG, Smith JA and Struhi K (1995) Current Protocols in Molecular Biology, supplement 36 (pp. 16.13.1–7) John Wiley & Sons, Inc.
Blasey HD, Aubry JP, Mazzei GJ and Bernard AR (1996) Large scale transient expression with COS cells. Cytotechnology 16: 183–192.
Cotter TG and Al-Rubeai M (1995) Cell death (apoptosis) in cell culture systems. Trends in BioTechnol 13: 150–155.
Fujita T, Terada S, Ueda H and Suzuki E (1996) Overexpression of bcl-2 improved survival of COS-1 cells and enhances transient protein production. J Fermentation and Bioengineering 82: 588–590.
Gauthier ER, Piche L, Lemieux G and Lemieux R (1996) Role of bcl-X(L) in the control of apoptosis in murine myeloma cells. Cancer Res. 56: 1451–1456.
Henderson S, Rowe M, Gregory C, Croom-Carter D, Wang F, Longnecker R, Kieff E and Rickinson A (1991) Induction of bcl-2 expression by Epstein-Barr virus latent membrane protein 1 products infected B cells from programmed cell death. Cell 65: 1107–1115.
Itoh Y, Ueda H and Suzuki E (1995) Overexpression of bcl-2, apoptosis suppressing gene: prolonged viable culture period of hybridoma and enhanced antibody production. Cytotechnology 48: 118–122.
Karasuyama H, Kudo A and Melchers F (1990) The proteins encoded by the VpreB and 5 pre-B cell-specific genes can associate with each other and with μ heavy chain. J Exp Med 172: 969–972.
Karasuyama H and Melchers F (1988) Establishment of mouse cell lines which constitutively secrete large quantities of interleukin 2, 3, 4 or 5, using modified cDNA expression vectors. Eur J Immunol 18: 97–104.
Mellon P, Parker V, Gluzmam Y and Maniatis T (1981) Identification of DNA sequences required for transcription of the human 1-globin gene in a new SV40 host-vector system. Cell 27: 279–288.
Miura M, Zhu H, Rotello R, Hartwieg EA and Yuan J (1993) Induction of apoptosis in fibroblasts by IL-1 beta-converting enzyme, a mammalian homologue of the C. elegans cell death gene ced-3. Cell 75: 653–660.
Murray K, Ang CE, Gull K, Hickman JA and Dickson JA (1996) NSO myeloma cell death: influence of bcl-2 overexpression. Biotechnol Bioeng 51: 298–304
Suzuki E, Terada S, Ueda H, Fujita T, Komatsu T, Kim YH, Takayama S and Reed JC (1997) Establishing apoptosis resistant cell lines for improving protein productivity of cell culture. Cytotechnology 23: 55–59.
Tsujimoto Y, Cossman J, Jaff E and Cruce CM (1985) Involvement of the bcl-2 gene in human follicular lymphoma. Science 228: 1440–1443.
Tsujimoto Y (1989) Overexpression of the human BCL-2 gene product results in growth enhancement of Epstein-Barr virusimmortalized B cells. Proc Natl Acad Sci USA 86: 1958–1962.
Tsujimoto Y (1989) Stress-resistance conferred by high level of bcl-2 protein in human B lymphoblastoid cell. Oncogene 4: 1331–1336.
Vaux DL, Cory S and Adams JM (1988) Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells. Nature 335: 440–442.
Williams GT and Smith CA (1993) Molecular regulation of apoptosis: genetic controls on cell death. Cell 74: 777–779.
Xue D and Horvitz R (1995) Inhibition of the Caenorhabditis elegans cell-death protease CED-3 by a CED-3 cleavage site in baculovirus p35 protein. Nature 377: 248–251.
Rights and permissions
About this article
Cite this article
Fujita, T., Terada, S., Fukuoka, K. et al. Reinforcing apoptosis-resistance of COS and myeloma cells by transfecting with bcl-2 gene. Cytotechnology 25, 25–33 (1997). https://doi.org/10.1023/A:1007935026770
Issue Date:
DOI: https://doi.org/10.1023/A:1007935026770