Heterologous protein expression by transimmortalized differentiated liver cell lines derived from transgenic mice (hepatomas/alpha 1 antitrypsin/ONC mouse).
A number of therapeutic plasma proteins are synthesized by human hepatocytes. Since many of these proteins undergo liver-specific post-translational modifications which are required for full biological activity, it may therefore be necessary to develop hepatocyte-based expression systems for their production. Using transgenic mice we have developed a transimmortalisation technique for the isolation of differentiated hepatic cell lines, already engineered to secrete human alpha 1 antitrypsin (alpha 1 AT), a plasma protein which is produced mainly in liver cells. This was achieved by co-expression of the mouse c-myc proto-oncogene and a genomic copy of the human alpha 1 AT gene, both under the control of the human alpha 1 AT promoter. Transgenic mice carrying this construct developed hepatomas producing human alpha 1 AT. Under defined culture conditions, cell lines secreting active alpha 1 AT were derived from these tumours. These cells maintain a differentiated hepatic phenotype and continue to secrete human alpha 1 AT for at least 40 generations.
Dalemans
W
W
Division of Molecular and Cellular Biology, Transgène S.A., Strasbourg, France.
Perraud
F
F
Le Meur
M
M
Gerlinger
P
P
Courtney
M
M
Pavirani
A
A
D000818:Animals; D001483:Base Sequence; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D003470:Culture Media; D016259:Genes, myc / Q000235:genetics; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational; D000090063:Proto-Oncogene Mas; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured / Q000378:metabolism*; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis* / Q000235:genetics
D003470:Culture Media; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D011994:Recombinant Proteins; D000515:alpha 1-Antitrypsin