Abstract
A novel two-compartment bioreactor, BelloCell®, was used to cultivate insect cells and a maximum yield of 4.6 × 109 cells was attained. The cells were immobilized in a packed bed fixed in the upper chamber, and the bellow in the lower chamber was compressed and released in an alternating fashion. The motion resulted in gentle, cyclic movement of the medium that was contained in the lower chamber and consequently exposed the cells to air in an oscillatory manner, thus rendering adequate aeration and uniform cell distribution in the bed. The baculovirus yield produced in BelloCell® could amount up to 3.3 × 1017 pfu using as little as 1.1 l medium in the production run. Besides, BelloCell® was extremely easy to handle and operate. These benefits underline the potential of BelloCell® for simple, economical and high-density cell culture and protein/virus production.
Similar content being viewed by others
References
Alauotila S., Marjamaki A., Matikainen M.T. and Jalkanen M. 1994. Use of a hollow-fiber bioreactor for large-scale production of alpha 2-adrenoceptors in mammalian cells. J. Biotechnol. 37: 179–184.
Chiou T.W., Wang Y.C. and Liu H.S. 1998. Utilizing the macroporous packed bed for insect cell/baculovirus expression. Part 2: The production of human interleukin-5 in polyurethane foam and cellulose foam packed bed bioreactors. Bioprocess Eng. 18: 91–100.
Chung I.S., Taticek R.A. and Shuler M.L. 1993. Production of human alkaline-phosphatase, a secreted, glycosylated protein, from a baculovirus expression system and the attachmentdependentcell-line Trichoplusia-Ni Bti-Tn 5B1-4 using a split-flow, airlift bioreactor. Biotechnol. Prog. 9: 675–678.
Hu Y.C. and Bentley W.E. 2000. A kinetic and statisticalthermodynamic model for baculovirus infection and viruslike particle assembly in suspended insect cells. Chem Eng Sci 55: 3991–4008.
Hu Y.C., Kaufman J., Cho M.W., Golding H. and Shiloach J. 2000. Production of HIV-1 gp120 in packed-bed bioreactor using the vaccinia virus/T7 expression system. Biotechnol. Prog. 16: 744–750.
Hu Y.C., Liu H.J. and Chung Y.C. 2002. High level expression of the key antigenic protein, σC, from avian reovirus into insect cells and its purification by immobilized metal affinity chromatography. Biotechnol. Lett. 24: 1017–1022.
Hu Y.C., Tsai C.T., Chung Y.C., Lu J.T. and Hsu J.T.A. 2003. Generation of chimeric baculovirus with histidine-tags displayed on the envelope and its purification using immobilized metal affinity chromatography. Enzyme Microb. Technol. 33: 445–452.
Ikonomou L., Drugmand J.C., Bastin G., Schneider Y.J. and Agathos S.N. 2002. Microcarrier culture of lepidopteran cell lines: Implications for growth and recombinant protein production. Biotechnol. Prog. 18: 1345–1355.
Kaufman J.B., Wang G., Zhang W., Valle M.A. and Shiloach J. 2000. Continuous production and recovery of recombinant Ca2+ binding receptor from HEK 293 cells using perfusion through a packed bed bioreactor. Cytotechnology 33: 3–11.
Kim Y., Sah R., Doong J. and Grodzinsky A. 1988. Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal. Biochem. 174: 168–176.
Kompier R., Kislev N., Segal I. and Kadouri A. 1991. Use of a stationary bed reactor and serum-free medium for the production of recombinant proteins in insect cells. Enzyme Microb. Technol. 13: 822–827.
Ma N., Koelling K.W. and Chalmers J.J. 2002. Fabrication and use of a transient contractional flow device to quantify the sensitivity of mammalian and insect cells to hydrodynamic forces. Biotechnol. Bioeng. 80: 428–436.
Obradovic B., Carrier R.L., Vunjak-Novakovic G. and Freed L.E. 1999. Gas exchange is essential for bioreactor cultivation of tissue engineered cartilage. Biotechnol. Bioeng. 63: 197–205.
O'Reilly D., Miller L. and Luckow V. 1992. Baculovirus expression vectors: a laboratory manual. W.H. Freeman and Co, New York, pp. 109–215.
Reiter M., Borth N., Blüml G., Wimmer K., Harant H., Zach N., Gaida T., Schmatz C. and Katinger H. 1992. Flow-cytometry and 2-dimensional electrophoresis (2-DE) for system evaluation of long term continuous perfused animal cell cultures in macroporous beads. Cytotechnology 9: 247–253.
Singh V. 1999. Disposable bioreactor for cell culture using waveinduced agitation. Cytotechnology 30: 149–158.
Tani H., Nishijima M., Ushijima H., Miyamura T. and Matsuura Y. 2001. Characterization of cell-surface determinants important for baculovirus infection. Virology 279:343–353.
Taticek R.A., Hammer D.A. and Shuler M.L. 1995. Overview of issues in bioreactor design and scale-up. In: Shuler M.L. (ed.), Baculovirus Expression Systems and Biopesticides, Wiley-Liss, New York, pp. 131–174.
Taticek R.A. and Shuler M.L. 1997. Effect of elevated oxygen and glutamine levels on foreign protein production at high cell densities using the insect cell-baculovirus expression system. Biotechnol. Bioeng. 54: 142–152.
Wickham T.J. and Nemerow G.R. 1993. Optimization of growth methods and recombinant protein production in Bti-Tn-5B1-4 insect cells using the baculovirus expression system. Biotechnol. Prog. 9: 25–30.
Wu J., King G., Daugulis A.J., Faulkner P., Bone D.H. and Goosen M.F.A. 1989. Engineering aspects of insect cell suspension culture: A review. Appl. Microbiol. Biotechnol. 32: 249–255.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hu, YC., Lu, JT. & Chung, YC. High-density cultivation of insect cells and production of recombinant baculovirus using a novel oscillating bioreactor. Cytotechnology 42, 145–153 (2003). https://doi.org/10.1023/B:CYTO.0000015841.98225.27
Issue Date:
DOI: https://doi.org/10.1023/B:CYTO.0000015841.98225.27