Chinese hamster ovary cells produce sufficient recombinant insulin-like growth factor I to support growth in serum-free medium - Serum-free growth of IGF-I producing CHO cells
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作者:
Hunt, SMN
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机构:UNIV NEW S WALES,DEPT BIOTECHNOL,KENSINGTON,NSW 2033,AUSTRALIA
Hunt, SMN
Pak, SCO
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机构:UNIV NEW S WALES,DEPT BIOTECHNOL,KENSINGTON,NSW 2033,AUSTRALIA
Pak, SCO
Bridges, MW
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机构:UNIV NEW S WALES,DEPT BIOTECHNOL,KENSINGTON,NSW 2033,AUSTRALIA
Bridges, MW
Gray, PP
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机构:UNIV NEW S WALES,DEPT BIOTECHNOL,KENSINGTON,NSW 2033,AUSTRALIA
Gray, PP
Sleigh, MJ
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机构:UNIV NEW S WALES,DEPT BIOTECHNOL,KENSINGTON,NSW 2033,AUSTRALIA
Sleigh, MJ
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[1] UNIV NEW S WALES,DEPT BIOTECHNOL,KENSINGTON,NSW 2033,AUSTRALIA
Insulin-like growth factor I has similar mitogenic effects to insulin, a growth factor required by most cells in culture, and it can replace insulin in serum-free formulations for some cells. Chinese Hamster Ovary cells grow well in serum-free medium with insulin and transferrin as the only exogenous growth factors. An alternative approach to addition of exogenous growth factors to serum-free medium is transfection of host cells with growth factor-encoding genes, permitting autocrine growth. Taking this approach, we constructed an IGF-I heterologous gene driven by the cytomegalovirus promoter, introduced it into Chinese Hamster Ovary cells and examined the growth characteristics of Insulin-like growth factor I-expressing clonal cells in the absence of the exogenous factor. The transfected cells secreted up to 500 ng/10(6) cells/day of mature Insulin-like growth factor I into the conditioned medium and as a result they grew autonomously in serum-free medium containing transferrin as the only added growth factor. This growth-stimulating effect, observed under both small and large scale culture conditions, was maximal since no further improvement was observed in the presence of exogenous insulin.