Characterization of recombinant human granulocyte-colony-stimulating factor produced in mouse cells.
Mouse C127I cells were transformed with a chimeric plasmid consisting of bovine papillomavirus DNA and human granulocyte-colony-stimulating factor (G-CSF) cDNA placed under the control of the SV40 early promoter. The transformed cells secreted constitutively a high level of human G-CSF, 10-20 micrograms/ml in a low-serum medium. The secreted G-CSF has been purified to homogeneity by a two-step procedure including gel filtration and hydrophobic column chromatography. The purified recombinant G-CSF runs as a single band with an apparent Mr of 19,000 on a polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. This value corresponds to that of the native human G-CSF purified from the medium conditioned by human carcinoma CHU-2 cells. The recombinant human G-CSF was as active as native G-CSF in vitro in supporting proliferation of mouse NFS-60 cells and stimulating colony formation from human as well as mouse bone marrow cells. When the recombinant human G-CSF was subcutaneously administrated into mice, a remarkable stimulation of granulopoiesis and splenomegaly was observed.
D000818:Animals; D002471:Cell Transformation, Neoplastic; D004247:DNA / Q000378:metabolism; D005796:Genes; D005814:Genes, Viral; D006410:Hematopoiesis / Q000187:drug effects*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D007377:Interleukin-3 / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D007700:Kinetics; D051379:Mice; D027383:Papillomaviridae / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D013539:Simian virus 40 / Q000235:genetics
D007377:Interleukin-3; D011994:Recombinant Proteins; D004247:DNA