, Volume 33, Issue 1–3, pp 37–46 | Cite as

Evaluation of stable and highly productive gene amplified CHO cell line based on the location of amplified genes

  • Tomohiro Yoshikawa
  • Fumi Nakanishi
  • Seima Itami
  • Daisuke Kameoka
  • Takeshi OmasaEmail author
  • Yoshio Katakura
  • Michimasa Kishimoto
  • Ken-ichi Suga


In order to establish an easy and quick construction method for obtaining a stable and highly productive gene-amplified recombinant Chinese Hamster Ovary (CHO) cell line, variouskinds of stepwise methotrexate (MTX) selection were carriedout. The specific growth and production rates of the cell were compared with each other, and the distribution of the amplified gene location was determined using fluorescence in situ hybridization (FISH). The specific growth andproduction rates of the cell pool reached the highest levels under the selection condition in which the stepwise increase in the MTX concentration was most gradual; about 82% of amplified genes were observed near the telomeric region. During long-term cultivation without MTX, the percentage ofamplified genes near the telomeric region hardly changed, butthat of amplified genes at other regions decreased. Based on these results, stable and highly productive cell pools could be easily and quickly constructed and amplified and gradual stepwise increase of the MTX concentration. In addition, the FISH technique was powerful tool to evaluate highly productiveand stable gene-amplified cells based on the chromosomal location of the amplified gene.

Chinese hamster ovary (CHO) dihydrofolate reductase (dhfr fluorescence in situ hybridization (FISH) gene amplification 


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Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Tomohiro Yoshikawa
    • 1
  • Fumi Nakanishi
    • 1
  • Seima Itami
    • 1
  • Daisuke Kameoka
    • 1
  • Takeshi Omasa
    • 2
    Email author
  • Yoshio Katakura
    • 1
  • Michimasa Kishimoto
    • 1
  • Ken-ichi Suga
    • 1
  1. 1.Department of Biotechnology, Graduate School of EngineeringOsaka UniversityOsakaJapan
  2. 2.Department of Biotechnology, Graduate School of EngineeringOsaka UniversityOsakaJapan

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