Screening of high-productivity cell lines and investigation of their physiology in Chinese hamster ovary (CHO) cell cultures for transforming growth factor-β1 production

A. Chun, A. Gie-Taek, Joo Buom Lee, Sang U. Nam, Se Won Lee, Yeon H. Jeong, Eui Yul Choi, Ik Hwan Kim, Yong Seob Jeong, Pyeong Hyeun Kim

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Using recombinant Chinese hamster ovary (CHO) cells, strategies for developing high producers for the recombinant human Transforming Growth Factor-β1 (TGF-β1) protein are proposed and their physiological characteristics in cell cultures were investigated. TGF-β1 is a pleiotrophic polypeptide involved in various biological activities, including cell growth, differentiation, and deposition of extracellular matrix proteins. The CHO cells included human TGF-β1 cDNA in conjunction with a dihydrofolate reductase (DHFR) gene, which was cotransfected into the cells to amplify the transfected TGF-β1 cDNA. As a first-round screening of the transfected cells, a relatively high TGF-β1-producing cell line was selected, and then, it acquired a resistance to increasing concentrations of methotrexate (MTX) up to 60 μM, resulting in a significant improvement in its TGF-β1 biosynthetic ability. After applying a monoclonal selection strategy to the MTX-resistant cells, more productive cells were screened, including the APP-3, App-5, and App-8 cell lines. These high producers were compared with two other cell lines (AP-1 cell line without amplification of transfected TGF-β1 cDNA and nontransfectant of TGF-β1 cDNA) in terms of cell growth, TGF-β1 productivity, sugar uptake, and byproduct formation, in the presence or absence of MTX in the culture medium. Consequently, both monoclonal selection as well as an investigation of the physiological characteristics were found to be needed for the efficient screening of higher TGF-β1 producers, even after the transfection and amplification of the transfected gene.

Original languageEnglish
Pages (from-to)121-129
Number of pages9
JournalJournal of Microbiology and Biotechnology
Volume12
Issue number1
Publication statusPublished - 2002 Jan 1
Externally publishedYes

Fingerprint

Physiology
Transforming Growth Factors
Cricetulus
Cell culture
Ovary
Screening
Cell Culture Techniques
Productivity
Cells
Cell Line
Complementary DNA
Methotrexate
Cell growth
Application programs
Amplification
Genes
Intercellular Signaling Peptides and Proteins
Proteins
Tetrahydrofolate Dehydrogenase
Gene Amplification

Keywords

  • Chinese hamster ovary (CHO) cells
  • CHO cell culture
  • Methotrexate (MTX)
  • Monoclonal selection
  • Transforming growth factor-β (TGF-β1)

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Screening of high-productivity cell lines and investigation of their physiology in Chinese hamster ovary (CHO) cell cultures for transforming growth factor-β1 production. / Chun, A.; Gie-Taek, A.; Lee, Joo Buom; Nam, Sang U.; Lee, Se Won; Jeong, Yeon H.; Choi, Eui Yul; Kim, Ik Hwan; Jeong, Yong Seob; Kim, Pyeong Hyeun.

In: Journal of Microbiology and Biotechnology, Vol. 12, No. 1, 01.01.2002, p. 121-129.

Research output: Contribution to journalArticle

Chun, A. ; Gie-Taek, A. ; Lee, Joo Buom ; Nam, Sang U. ; Lee, Se Won ; Jeong, Yeon H. ; Choi, Eui Yul ; Kim, Ik Hwan ; Jeong, Yong Seob ; Kim, Pyeong Hyeun. / Screening of high-productivity cell lines and investigation of their physiology in Chinese hamster ovary (CHO) cell cultures for transforming growth factor-β1 production. In: Journal of Microbiology and Biotechnology. 2002 ; Vol. 12, No. 1. pp. 121-129.
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abstract = "Using recombinant Chinese hamster ovary (CHO) cells, strategies for developing high producers for the recombinant human Transforming Growth Factor-β1 (TGF-β1) protein are proposed and their physiological characteristics in cell cultures were investigated. TGF-β1 is a pleiotrophic polypeptide involved in various biological activities, including cell growth, differentiation, and deposition of extracellular matrix proteins. The CHO cells included human TGF-β1 cDNA in conjunction with a dihydrofolate reductase (DHFR) gene, which was cotransfected into the cells to amplify the transfected TGF-β1 cDNA. As a first-round screening of the transfected cells, a relatively high TGF-β1-producing cell line was selected, and then, it acquired a resistance to increasing concentrations of methotrexate (MTX) up to 60 μM, resulting in a significant improvement in its TGF-β1 biosynthetic ability. After applying a monoclonal selection strategy to the MTX-resistant cells, more productive cells were screened, including the APP-3, App-5, and App-8 cell lines. These high producers were compared with two other cell lines (AP-1 cell line without amplification of transfected TGF-β1 cDNA and nontransfectant of TGF-β1 cDNA) in terms of cell growth, TGF-β1 productivity, sugar uptake, and byproduct formation, in the presence or absence of MTX in the culture medium. Consequently, both monoclonal selection as well as an investigation of the physiological characteristics were found to be needed for the efficient screening of higher TGF-β1 producers, even after the transfection and amplification of the transfected gene.",
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AU - Lee, Se Won

AU - Jeong, Yeon H.

AU - Choi, Eui Yul

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AU - Jeong, Yong Seob

AU - Kim, Pyeong Hyeun

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