Skip to main content
Log in

High-density cultivation of insect cells and production of recombinant baculovirus using a novel oscillating bioreactor

  • Published:
Cytotechnology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Google Scholar 

  • 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.

    PubMed  CAS  Google Scholar 

  • Singh V. 1999. Disposable bioreactor for cell culture using waveinduced agitation. Cytotechnology 30: 149–158.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yu-Chen Hu.

Rights and permissions

Reprints 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

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:CYTO.0000015841.98225.27

Navigation