Growth and secretion of erythropoietin of Chinese hamster ovary cells coexpressing epidermal growth factor receptor and erythropoietin genes: Design of cells for cell culture matrix.
Chinese hamster ovary (CHO) cells were transfected withboth genes encoding erythropoietin (Epo) and epidermal growthfactor receptor (EGFR). The transfection of the Epo gene wasconfirmed by an enzyme-linked immunoassay. Overexpression ofEGFR was confirmed by Western blotting of EGFR. Thetransfected CHO cells were cultured in serum-free medium inthe presence of soluble epidermal growth factor (EGF) orimmobilized EGF. The CHO cells overexpressing EGFR grew in thepresence of less EGF than the cells not overexpressing EGFR.In addition, the growth of EGFR-overexpressing CHO cells wasenhanced in the presence of immobilized EGF more efficientlythan in the presence of soluble EGF. The amount of Eposecreted from the cells increased linearly with the increaseof growth rate. Consequently, culture of CHO cellscoexpressing Epo and EGFR on EGF-immobilized matrix was themost efficient for Epo production.
Chen
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G
3D Tissue Engineering Group, National Institute for Advanced Interdisciplinary Research, 1-1-4 Higashi, Tsukuba, Ibaraki, 305-8562, Japan.
Ito
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Y
Masuda
S
S
Sasaki
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R