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Perfusion culture apparatus for suspended mammalian cells

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Abstract

A variety of processes have been proposed for mammalian cell culture in the commercial production of useful substances (e.g., monoclonal antibodies, therapeutic and diagnostics proteins). Among them, the perfusion culture of suspended non-immobilized cells is the most advantageous. Perfusion culture can be classified by the separation process of suspended cells from the culture mixture into three types, namely filtration, gravitational settling and centrifugation. From a commercial point of view, the present situation and technical problems of suspended-cell perfusion culture will be reviewed based on the three types, The recent development of perfusion culture has been carried out mainly on the filtration separation process, but the centrifugation process seems to have a promising future because of operation stability and scale-up feasibility. The reasons will be explained in details.

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References

  • Apelman S (1992) Separation of animal cells in continuous cell culture system. In: Murakami H, Shirahata S and Tachibana H (eds.) Animal Cell Technology: Basic and Applied Aspects (pp. 149–154) Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Arai T, Yokoyama S and Tokashiki M (1993) 50L Scale perfusion culture of hybridoma cells by gravitational settling for cell separation. In: Kaminogawa S, Ametani A and Hachimura S (eds.) Animal Cell Technology: Basic and Applied Aspects. (pp. 341–346) Kluwer Academic Publishers, Dordrecht.

    Google Scholar 

  • Avgerinos GC, Drapeau D, Socolow JS, Mao J, 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: 54–58.

    Google Scholar 

  • Blasey HD and Jäger V (1991) Strategies to increase the efficiency of membrane aerated and perfused animal cell bioreactors by an improved medium perfusion. In: Sasaki R and Ikura K (eds.) Animal Cell Culture Production of Biologicals. (pp. 61–73) Kluwer Academic Publishers, Dordrecht.

    Google Scholar 

  • Brennan AJ, Shevitz J and Macmillian J (1987) A perfusion system for antibody production by shear sensitive hybridoma cells in a stirred reactor. Biotechnol. Tech. 1: 169–174.

    Google Scholar 

  • Büntemeyer H, Lutkemeyer D and Lehmann J (1991) Optimization of serum-free fermentation processes for antibody production. Cytotechnology 5: 57–67.

    Google Scholar 

  • Hamamoto K, Tokashiki M and Ichikawa Y (1987) High cell density culture of a hybridoma using perfluorocarbon to supply oxygen. Agric. Biol. Chem. 51: 3415–3416.

    Google Scholar 

  • Hamamoto K, Ishimaru K and Tokashiki M (1989) Perfusion culture of hybridoma cells using a centrifuge to separate cells from culture mixture. J. Ferment. Bioeng. 67: 190–194.

    Google Scholar 

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

    Google Scholar 

  • Hosoi S, Miyaji H, Satoh M, Kurimoto T, Mihara A, Fujiyoshi H, Itoh S and Sato S (1991a) Optimization of cell culture conditions for production of biologically active proteins. Cytotechnology 5: S17–34.

    Google Scholar 

  • Hosoi S, Murosumi K, Sasaki K, Satoh M, Miyaji H, Hasegawa M, Itoh S, Tamaoki T and Sato S (1991b) Optimization of cell culture conditions for G-CSF (granulocyte-colony stimulating factor) production by genetically engineered Namalwa KJM-1 cells. Cytotechnology 7: 25–32.

    Google Scholar 

  • Ishida M, Haga R, Matsuzaki H and Nakano R (1990) High cell density suspension culture of mammalian anchorage-independent cells: Oxygen transfer by gas sparging and deforming with hydrophobic net. Cytotechnology 4: 215–225.

    Google Scholar 

  • Jäger V (1992) High density perfusion culture of animal cells using a novel continuous flow centrifuge. In: Murakami H, Shirahata S and Tachibana H (eds.) Animal Cell Technology: Basic and Applied Aspects. (pp. 209–216) Kluwer Academic Publishers, Dordrecht.

    Google Scholar 

  • Kitano K, Shintani Y, Ichimori Y, Tsukamoto K, Sasai S and Kida M, (1986) Production of human monoclonal antibodies by heterohybridomas. Appl. Microbiol. Biotechnol. 24: 282–286.

    Google Scholar 

  • Kitano K, Ichimori Y, Sawada H, Iwasai S and Tsukamoto K (1991) Effective production of anti-tetanus toxoid and anti-HBsAg human monoclonal antibodies by serum-free culture of hybridomas. Cytotechnology 5: S53–74.

    Google Scholar 

  • Lehmann J, Vorlop J and Büntemeyer H (1988) Bubble-free reactors and their development for continuous culture with cell recycle. In: Spier RE and Griffiths JB (eds.) Animal Cell Biotechnology. vol 3 (pp. 221–237) Academic Press Ltd., London.

    Google Scholar 

  • Matsumura M, Misato T, Kataoka H and Mayumi M (1989) Production of HBs-Mabs by perfusion culture in membrane reactor. In: Murakami H (ed.) Trends in Animal Cell Technology. (pp. 145–148) Kodansha Ltd.

  • Murakami H, Shimomura T, Ohashi H, Hashizume S, Tokashiki M, Shinohara K, Yasumoto K, Nomoto K and Omura H (1985) Serum free stirred culture of human-human hybridoma lines. In: Growth and Differentiation of Cells in Defined Environment. (pp. 111–116) Kodansha Ltd. and Springer-Verlag.

  • Nelson KL (1988) Industrial-scale mammalian cell culture, part 1: Bioreactor design considerations. Biopharm. Manufact. Feb. 42–46.

  • Phillips AW, Ball GD, Fantes KH, Finter NB and Johnson MD (1985) Experience in the cultivation of mammalian cells on the 8000 1 scale. In: Feder J and Tolbert WR (eds.) Large-Scale Mammalian Cell Culture. (pp. 87–95) Academic Press, Inc., Orlando.

    Google Scholar 

  • Pullen KF, Jonson MD, Philips AW, Ball GD and Finter NB (1985) Very large scale suspension cultures of mammalian cells. Develop. Biol. Standard. 60: 175–177.

    Google Scholar 

  • Rebsamen E, Goldinger W, Merten OW and Palfi GE (1987). Use of a dynamic filtration method for separation of animal cells. Develop. Biol. Standard. 66: 273–277.

    Google Scholar 

  • Reuveny S, Velez D, Macmillan JD and Miller L (1986) Factors affecting cell growth and monoclonal antibody production in stirred reactors. J. Immunologic. Meth. 86: 53–59.

    Google Scholar 

  • Sato S, Kawamura K and Fujiyoshi N (1983a) Mass production of animal cells in middle scale—interferon production by high density cell culture. Soshiki Baiyo. 8: 286–290.

    Google Scholar 

  • Sato S, Kawamura K and Fujiyoshi N (1983b) Animal cell cultivation for production of biological substances with a novel perfusion culture apparatus.J. Tissue Culture Methods 8: 167–171.

    Google Scholar 

  • Sato S, Kawamura K, Hanai N and Fujiyoshi N (1985) Production of interferon and monoclonal antibody using a novel type of perfusion vessel. In: Murakami H, Yamane I, Barnes DW, Mather JP, Hayashi I and Sato GH (eds.) Growth and Differentiation of Cell in Defined Environment. (pp. 123–126) Kodansha Ltd., Tokyo and Springer-Verlag, Heidelberg.

    Google Scholar 

  • Seamans TC and Hu WS (1990) Kinetics of growth and antibody production by a cell line in a perfusion culture. J. Ferment. Biotech. 70: 535–536.

    Google Scholar 

  • Shimoda M, Matsumura M and Kataoka H (1991) Growth characteristics of hybridoma cells under glucose limiting conditions. Kagaku Kogaku Ronbunshu 17: 642–648.

    Google Scholar 

  • Shintani Y, Kohno Y, Sawada H and Kitano K (1991) Comparison of culture methods for human-human hybridomas secreting anti-HBsAg human monoclonal antibodies. Cytotechnology 6: 197–208.

    Google Scholar 

  • Takazawa Y, Tokashiki M, Murakami H, Yamada K and Omura H (1988) High-density culture of mouse-mouse hybridoma in serum-free defined medium. Biotechnol. Bioeng. 31: 168–172.

    Google Scholar 

  • Takazawa Y and Tokashiki M (1989a) High cell density perfusion culture of mouse-human hybridomas. Appl. Microbiol. Biotechnol. 32: 280–284.

    Google Scholar 

  • Takazawa Y and Tokashiki M (1989b) Production of human-mouse chimeric antibody by high cell density perfusion culture. Cytotechnology 2: 95–101.

    Google Scholar 

  • Tokashiki M, Takazawa Y and Hamamoto K (1987) High density culture of hybridoma cells using a perfusion culture vessel with filter. Hakkokogaku 65: 535–536.

    Google Scholar 

  • Tokashiki M, Hamamoto K, Takazawa Y and Ichikawa Y (1988) High-density culture of mouse-human hybridoma cells using a new perfusion culture vessel. Kagakukogaku Ronbunshu 14: 337–341.

    Google Scholar 

  • Tokashiki M and Arai T (1989) High density culture of mouse-human hybridoma cells using a perfusion culture apparatus with multi-settling zones to separate cells from the culture medium. Cytotechnology 2: 5–8.

    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.

    Google Scholar 

  • Tokashiki M and Arai T (1991) In: Spier RE, Griffiths JB and Meignier B (eds.) Production of Biologicals from Animal Cells in Culture. (pp. 467–469) Butterworth-Heinemann, Oxford.

    Google Scholar 

  • Tolbert WR, Feder J and Kimers RS (1981) Large scale rotating filter perfusion system for high density growth of mammalian suspension cultures. In Vitro 17: 885–890.

    Google Scholar 

  • Tolbert WR and Feder J (1983) Large-scale cell culture technology. In: Tsao GT, Flickinger MC and Finn RK (eds.) Annual Reports on Fermentation Processes 6. (pp. 35–74) Academic Press, Inc., New York.

    Google Scholar 

  • Tolbert WR, Lewis C, White PJ and Feder J (1985) In: Feder J and Tolbert WR (eds.) Large-Scale Mammalian Cell Culture. (pp. 97–123) Academic Press, Inc., Orlando.

    Google Scholar 

  • Tolbert WR, Srigley WR and Prior CP (1988) In: Spier RE and Griffiths JB (eds.) Animal Cell Biotechnology, Vol. 3 (pp. 373–393) Academic Press Ltd., London.

    Google Scholar 

  • Van Wezel AL, van der Velden-de Groot CAM, de Haan JJ, van den Heuval N and Schasfoort R (1985) Large scale animal cell cultivation for production of cellular biologicals. Dev. Biol. Stand. 60: 229–236.

    Google Scholar 

  • Velez D, Reuveny S, Miller L and Macmillan JD (1987) Effect of feeding rate on monoclonal antibody production in a modified perfusion-fed fermentor. J. Immunologic. Meth. 102: 275–278.

    Google Scholar 

  • Velez D, Miller L and Macmillan JD (1989) Use of tangential flow filtration in perfusion propagation of hybridoma cells for propagation of monoclonal antibodies. Biotechnol. Bioeng. 33: 938–940.

    Google Scholar 

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Tokashiki, M., Takamatsu, H. Perfusion culture apparatus for suspended mammalian cells. Cytotechnology 13, 149–159 (1993). https://doi.org/10.1007/BF00749811

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