Cytotechnology

, Volume 14, Issue 1, pp 61–66 | Cite as

High-density culture of FM-3A cells using a bioreactor with an external tangential-flow filtration device

  • Hiroyuki Kawahara
  • Shinjiro Mitsuda
  • Eitaro Kumazawa
  • Yasuyoshi Takeshita
Article

Abstract

A novel bioreactor system developed for high-density cultures of suspended mammalian cells is described using a tangential-flow filtration device outside the culture vessel to separate viable cells from spent medium. The filtration device is based on thin porous microfiltration membranes with a pore size of 0.20–0.65 μm. Because cells have a diameter of about 10–20 μm, they cannot permeate these membranes with the spent medium. So, allowing a perfusion culture to be created using this system. In most membrane filtration systems, clogging of the membranes has made long-term operation difficult. In this system, however, high pressure is not applied directly to the membrane, thus minimizing clogging. Also, clogging of the membrane was prevented by washing the membrane surface once a day, and increasing the membrane surface are. With this system, FM-3A cells were cultured and maintained at a high density of 3.0×107 cells/ml for two weeks, and a continuous culture was supported for as long as 34 days.

Key words

FM-3A cells high-density culture perfusion culture tangential-flow filtration 

Abbreviation

DO

dissolved oxygen

PVDF

polyvinylidene di-fluoride

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Broise D, Noiseux M, Massie B and Lemieux R (1992) Hybridoma perfusion systems: A comparison study. Biotech. Bioeng. 40: 25–32.Google Scholar
  2. Fraune E (1989) Filtration systems for the perfusion of cell culture media. Paper presented at the 1st Taunus Conference Membrane Technology, 3/89 Bad Soden, West-Germany.Google Scholar
  3. Himmelfarb P, Thayer PS and Martin HE (1969) Spin filter culture: The propagation of cells in suspension. Science 164: 555–557.Google Scholar
  4. Knazek PA, Gullino PM, Kohler PO and Dedrick RL (1972) Cell culture on artificial capillaries. Science 178: 65–67.Google Scholar
  5. Koyama H (1980) FM-3A cell line. Soshiki Baiyou 6(7): 235–243.Google Scholar
  6. Lim F and Moss RD (1981) Microencapsulation of living cells and tissues. J. Pharm. Sci. 70: 351–354.Google Scholar
  7. Nilsson K, Scheirer W, Merten OW, Osteberg L, Liehl E, Katinger HWD and Mosbach K (1983) Entrapment of animal cells for production of monoclonal antibodies and other biochemicals. Nature 302: 629–630.Google Scholar
  8. Satoh S, Kawamura K and Fujiyoshi N (1983) Animal cell cultivation of biological substances with a novel perfusion culture apparatus. J. Tissue Culture Methods 8: 167–171.Google Scholar
  9. Seamans CT and Hu W-S (1990) Kinetics of growth and antibody production by hybridoma cell line in a perfusion culture. J. Ferm. Bioeng. 70: 241–245.Google Scholar
  10. Tokashiki M, Hamamoto K, Takazawa Y and Ichikawa Y (1988) High-density culture of mouse-human hybridoma cells using a new perfusion culture vessel. Kagaku Kogaku Ronbunshu 14: 337–341.Google Scholar
  11. 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
  12. Velez D, Miller L and Macmillan DJ (1989) Use of tangential flow filtration in perfusion propagation of hybridoma cells for production of monoclonal antibodies. Biotech. Bioeng. 33: 938–940.Google Scholar

Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Hiroyuki Kawahara
    • 1
  • Shinjiro Mitsuda
    • 1
  • Eitaro Kumazawa
    • 1
  • Yasuyoshi Takeshita
    • 1
  1. 1.Research Institute of Life ScienceSnow Brand Milk Products Co., Ltd.TochigiJapan

Personalised recommendations