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Cytotechnology

, Volume 6, Issue 2, pp 93–103 | Cite as

Improvement in the proliferative activity of human-human hybridomas at low cell density by transfection with bFGF gene

  • Keiji Iwamoto
  • Yasushi Shintani
  • Hidekazu Sawada
  • Kazuaki Kitano
Original Research Paper

Abstract

Highly purified recombiaant basic fibroblast growth factor (rbFGF) and acidic FGF (aFGF) stimulated the proliferation of human-human (h-h) hybridomas to the extent of over four-fold from a low cell density such as 1×103 cells per ml in a serum-free medium in 24-well plates. The stimulatory effect of rbFGF was also observed in various lymphoid cell lines. Expecting that FGF could be an autocrine growth factor, we introduced bFGF gene into a h-h hybridoma using an expression plasmid induced by dexamethasone. The transformed cells thus obtained, HPO-75.11bbFGF-7, were able to grow well from a low inoculum density in a serum-free medium and antibody production was also increased when bFGF gene expression was induced. The transformed cells could grow at clonal density in a serum-free medium in 96-well plates, though the original cells could not. We also obtained a more practical transfectant, HPO-75.29-H74, using a high-shear stress adapted clone as the recipient and an expression plasmid having bFGF gene under the control of metallothioneine-I promoter. The HOP-75.29-H74 cells were capable of growing and producing human monoclonal antibody against hepatitis B virus surface antigen from an inoculum density of 1×103 cells per ml in an agitation vessel without addition of an inducer.

Key words

cell improvement FGF hepatitis B virus human-human hybridoma monoclonal antibody proliferative activity 

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Copyright information

© Kluwer Academic Publishers 1991

Authors and Affiliations

  • Keiji Iwamoto
    • 1
  • Yasushi Shintani
    • 1
  • Hidekazu Sawada
    • 1
  • Kazuaki Kitano
    • 1
  1. 1.Microbiology Research Laboratories, Research and Development DivisionTakeda Chemical Industries, Ltd.OsakaJapan

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