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Long-term stable production of monocyte-colony inhibition factor (M-CIF) from CHO microcarrier perfusion cultures

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Abstract

Monocyte-colony inhibition factor (M-CIF) was produced in microcarrier perfusion cultures from engineered Chinese hamster ovary (CHO) cells. Three and fifteen liter microcarrier perfusion bioreactors equipped with internal spin filters were operated for over two months. Approximately 60 L and 300 L of culture filtrate were harvested from the 3L and 15L microcarrier perfusion bioreactors respectively. During the perfusion operation, cell density reached 2–6 × 106 cells/ml. Importantly, stable expression of M-CIF from the CHO cells under non-selection condition was maintained at a level of 4–10 mg/L. Specific productivity was maintained at 1.8–3.4 mg/billion cells/day. The ability of the recombinant CHO cells to migrate from microcarrier to microcarrier under our proprietary HGS-CHO-3 medium greatly facilitated microcarrier culture scale-up and microcarrier replenishment. Future directions for microcarrier perfusion system scale-up and process development are highlighted.

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References

  • Avgerinos GC, Drapeau D, Socolow JS, Mao JI, Hsiao K and Broeze RJ (1990) Spin filter perfusion system for high density cell culture: Production of recombinant urinary type plasminogen activator in CHO cells. Bio/Technology 8(1): 54-58.

    Article  CAS  Google Scholar 

  • Banik GG and Heath CA (1995) Partial and total cell retention in a filtration-based homogeneous perfusion reactor. Biotechnol Prog 11(5): 584-588.

    Article  CAS  Google Scholar 

  • Cartwright T (1994) Introducing the animal cell as a bioreactor. In: Cartwright T (ed.) Animal Cells as Bioreactors. (pp. 1-10) Cambridge University Press, Cambridge, UK.

    Google Scholar 

  • Emery AN, Jan DCH and al-Rubeai M(1995) Oxygenation of intensive cell-culture system. Appl Microbiol Biotechnol 43: 1028-33.

    Article  CAS  Google Scholar 

  • Fenge C, Klein C, Heuer C, Siegel U and Fraune E (1993) Agitation, aeration and perfusion modules for cell culture bioreactors. Cytotechnology 11(3): 233-244.

    Article  CAS  Google Scholar 

  • Griffiths JB (1988) Overview of cell culture systems and their scale-up. In: Spier RE and Griffiths JB (eds.) Animal Cell Biotechnology, Vol. 3(pp. 179-220) Academic Press, London.

    Google Scholar 

  • Hansen HA, Damgaard B and Emborg C (1993) Enhanced antibody production associated with altered amino acid metabolism in a hybridoma high-density perfusion culture established by gravity separation. Cytotechnology 11(2): 155-166.

    Article  CAS  Google Scholar 

  • Hayter PM, Curling EM, Baines AJ, Jenkins N, Salmon I, Strange PG and Bull AT (1991) Chinese hamster ovary cell growth and interferon production kinetics in stirred batch culture. Appl Microbiol Biotechnol 34: 559-564.

    Article  CAS  Google Scholar 

  • Himmelfarb P, Thayer PS and Martin HE (1969) Spin filter culture: The propagation of mammalian cells in suspension. Science 164: 555-557.

    CAS  Google Scholar 

  • Kreider BL, Zhang J, Li H, Su J, Oelkuct M, Gentz R, Garotta G and Patel VP (1996) A beta-family chemokine which specifically inhibits MCSF mediated colony formation. Eur Cytokine Netw 7(3): 493.

    Google Scholar 

  • Miller AO, Menozzi FD and Dubois D (1989) Microbeads and anchorage-dependent eukaryotic cells: the beginning of a new era in biotechnology. Adv Biochem Eng Biotechnol 39: 73-95.

    Google Scholar 

  • Nilsson K(1989) Microcarrier cell culture. Biotechnol. Genetic Eng. Rev. 6 (September): 403-439.

  • Ogata M, Wakita K, Kimura K, Marumoto Y, Ohi K and Shimizu S (1993) High-level expression of recombinant human soluble thrombomodulin in serum-free medium by CHOK1 cells. Appl Microbiol Biotechnol 38(4): 520-5.

    Article  CAS  Google Scholar 

  • Omasa T, Kobayashi M, Nishikawa T, Shioya S, Suga KI, Uemura SI, Kitani Y and Imamura Y (1995) Enhancement of antibody production by growth factor addition in perfusion and hollow-fiber culture systems. Biotechnol Bioeng 48: 673-680.

    Article  CAS  Google Scholar 

  • Ross, SM (1987) Introduction to Probability and Statistics for Engineers and Scientists. John Wiley & Sons, New York.

    Google Scholar 

  • Searles JA, Todd P and Kompala DS (1994) Viable cell recycle with an inclined settler in the perfusion culture of suspended recombinant Chinese hamster ovary cells. Biotechnol Prog 10(2): 198-206.

    Article  CAS  Google Scholar 

  • Tokashiki M, Arai T, Hamamoto K and Ishimaru K (1990) High density culture of hybridoma cells using a perfusion culture vessel with an external centrifuge. Cytotechnology 3: 239-244.

    Article  CAS  Google Scholar 

  • Urlaub G, Kas E, Carothers AM and Chasin LA (1983) Deletion of the diploid dihydrofolate reductase locus from cultured mammalian cells. Cell 33(2): 405-412.

    Article  CAS  Google Scholar 

  • Urlaub G, Mitchell PJ, Kas E, Chasin LA, Funanage VL, Myoda TT and Hamlin J (1986) Effect of gamma rays at the dihydrofolate reductase locus: Deletions and inversions. Somat Cell Mol Genet 12(6): 555-566.

    Article  CAS  Google Scholar 

  • Zhang J, Kreider BL, Li H, Su J, Zhang JL, Gentz R, Garotta G and Antonaceio M (1996) Preventative effect of a β-chemokine on LPS-induced septic shock. Eur Cytokine Netw 7(3): 207.

    Google Scholar 

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Kong, D., Gentz, R. & Zhang, J. Long-term stable production of monocyte-colony inhibition factor (M-CIF) from CHO microcarrier perfusion cultures. Cytotechnology 26, 131–138 (1998). https://doi.org/10.1023/A:1007997412002

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  • DOI: https://doi.org/10.1023/A:1007997412002

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