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Characterization of recombinant human granulocyte‐colony‐stimulating factor produced in mouse cells.
Author(s) -
Tsuchiya M.,
Nomura H.,
Asano S.,
Kaziro Y.,
Nagata S.
Publication year - 1987
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1987.tb04798.x
Subject(s) - biology , recombinant dna , granulocyte , granulocyte colony stimulating factor , microbiology and biotechnology , virology , immunology , genetics , gene , chemotherapy
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.