Skip to main content
Cytotechnology logoLink to Cytotechnology
. 1998 Mar;26(2):131–138. doi: 10.1023/A:1007997412002

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

Deyu Kong 1, Reiner Gentz 1, Junli Zhang 1
PMCID: PMC3466677  PMID: 22358551

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.

Keywords: CHO, microcarrier, M-CIF, perfusion, spin filter, stable production

Full Text

The Full Text of this article is available as a PDF (174.4 KB).

References

  1. Avgerinos GC, Drapeau D, Socolow JS, Mao JI, Hsiao K, Broeze RJ. Spin filter perfusion system for high density cell culture: Production of recombinant urinary type plasminogen activator in CHO cells. Bio/Technology. 1990;8(1):54–58. doi: 10.1038/nbt0190-54. [DOI] [PubMed] [Google Scholar]
  2. Banik GG, Heath CA. Partial and total cell retention in a filtration-based homogeneous perfusion reactor. Biotechnol Prog. 1995;11(5):584–588. doi: 10.1021/bp00035a013. [DOI] [PubMed] [Google Scholar]
  3. Cartwright T. Introducing the animal cell as a bioreactor. In: Cartwright T, editor. Animal Cells as Bioreactors. Cambridge, UK: Cambridge University Press; 1994. pp. 1–10. [Google Scholar]
  4. Emery AN, Jan DCH, al-Rubeai M. Oxygenation of intensive cell-culture system. Appl Microbiol Biotechnol. 1995;43:1028–33. doi: 10.1007/BF00166920. [DOI] [PubMed] [Google Scholar]
  5. Fenge C, Klein C, Heuer C, Siegel U, Fraune E. Agitation, aeration and perfusion modules for cell culture bioreactors. Cytotechnology. 1993;11(3):233–244. doi: 10.1007/BF00749874. [DOI] [PubMed] [Google Scholar]
  6. Griffiths JB. Overview of cell culture systems and their scale-up. In: Spier RE, Griffiths JB, editors. Animal Cell Biotechnology. London: Academic Press; 1988. pp. 179–220. [Google Scholar]
  7. Hansen HA, Damgaard B, Emborg C. Enhanced antibody production associated with altered amino acid metabolism in a hybridoma high-density perfusion culture established by gravity separation. Cytotechnology. 1993;11(2):155–166. doi: 10.1007/BF00749005. [DOI] [PubMed] [Google Scholar]
  8. Hayter PM, Curling EM, Baines AJ, Jenkins N, Salmon I, Strange PG, Bull AT. Chinese hamster ovary cell growth and interferon production kinetics in stirred batch culture. Appl Microbiol Biotechnol. 1991;34:559–564. doi: 10.1007/BF00167898. [DOI] [PubMed] [Google Scholar]
  9. Himmelfarb P, Thayer PS, Martin HE. Spin filter culture: The propagation of mammalian cells in suspension. Science. 1969;164:555–557. doi: 10.1126/science.164.3879.555. [DOI] [PubMed] [Google Scholar]
  10. Kreider BL, Zhang J, Li H, Su J, Oelkuct M, Gentz R, Garotta G, Patel VP. A beta-family chemokine which specifically inhibits MCSF mediated colony formation. Eur Cytokine Netw. 1996;7(3):493. [Google Scholar]
  11. Miller AO, Menozzi FD, Dubois D. Microbeads and anchorage-dependent eukaryotic cells: the beginning of a new era in biotechnology. Adv Biochem Eng Biotechnol. 1989;39:73–95. doi: 10.1007/BFb0051952. [DOI] [PubMed] [Google Scholar]
  12. Nilsson K(1989) Microcarrier cell culture. Biotechnol. Genetic Eng. Rev. 6 (September): 403-439. [PubMed]
  13. Ogata M, Wakita K, Kimura K, Marumoto Y, Ohi K, Shimizu S. High-level expression of recombinant human soluble thrombomodulin in serum-free medium by CHOK1 cells. Appl Microbiol Biotechnol. 1993;38(4):520–5. doi: 10.1007/BF00242948. [DOI] [PubMed] [Google Scholar]
  14. Omasa T, Kobayashi M, Nishikawa T, Shioya S, Suga KI, Uemura SI, Kitani Y, Imamura Y. Enhancement of antibody production by growth factor addition in perfusion and hollow-fiber culture systems. Biotechnol Bioeng. 1995;48:673–680. doi: 10.1002/bit.260480616. [DOI] [PubMed] [Google Scholar]
  15. Ross SM. Introduction to Probability and Statistics for Engineers and Scientists. New York: John Wiley & Sons; 1987. [Google Scholar]
  16. Searles JA, Todd P, Kompala DS. Viable cell recycle with an inclined settler in the perfusion culture of suspended recombinant Chinese hamster ovary cells. Biotechnol Prog. 1994;10(2):198–206. doi: 10.1021/bp00026a600. [DOI] [PubMed] [Google Scholar]
  17. Tokashiki M, Arai T, Hamamoto K, Ishimaru K. High density culture of hybridoma cells using a perfusion culture vessel with an external centrifuge. Cytotechnology. 1990;3:239–244. doi: 10.1007/BF00365487. [DOI] [PubMed] [Google Scholar]
  18. Urlaub G, Kas E, Carothers AM, Chasin LA. Deletion of the diploid dihydrofolate reductase locus from cultured mammalian cells. Cell. 1983;33(2):405–412. doi: 10.1016/0092-8674(83)90422-1. [DOI] [PubMed] [Google Scholar]
  19. Urlaub G, Mitchell PJ, Kas E, Chasin LA, Funanage VL, Myoda TT, Hamlin J. Effect of gamma rays at the dihydrofolate reductase locus: Deletions and inversions. Somat Cell Mol Genet. 1986;12(6):555–566. doi: 10.1007/BF01671941. [DOI] [PubMed] [Google Scholar]
  20. Zhang J, Kreider BL, Li H, Su J, Zhang JL, Gentz R, Garotta G, Antonaceio M. Preventative effect of a β-chemokine on LPS-induced septic shock. Eur Cytokine Netw. 1996;7(3):207. [Google Scholar]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.

RESOURCES