Maintenance of liver function in long term culture of hepatocytes following in vitro or in vivo Ha-rasEJ transfection
| Autor: | Michel Dreano, Frederic Montandon, Hanwei Cao, Mohammed El Baraka, Miguel Diez Ibanez, Christos Tsaconas, Sami Alouani, Martine Chessebeuf-Padieu, Prudent Padieu, Michel Fischbach |
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| Rok vydání: | 1991 |
| Předmět: |
Male
medicine.medical_specialty Health Toxicology and Mutagenesis Blotting Western Gene Expression Biology Toxicology Transfection Cell Line Bile Acids and Salts In vivo Internal medicine Albumins medicine Animals Cells Cultured Cell Line Transformed chemistry.chemical_classification Albumin Transferrin Rats Inbred Strains Cell Biology 2-Acetylaminofluorene Molecular biology In vitro Rats Blotting Southern Endocrinology medicine.anatomical_structure Genes ras chemistry Liver Cell culture Hepatocyte Growth Hormone Liver function Cell Division |
| Zdroj: | Cell biology and toxicology. 7(4) |
| ISSN: | 0742-2091 |
| Popis: | Collagenase isolated rat hepatocytes were transfected with liposome encapsulated pEJ (LE-pEJ), a plasmid carrying the human cellular activated Ha-rasEJ oncogene. A proliferative cell line was cloned from these cells transfected in vitro. It secreted per day 0.87 micrograms albumin and 0.32 microgram transferrin per 10(6) cells, and 11.06 nmol free and conjugated bile acids (BA) per mg protein. Also, it metabolized 2-acetylaminofluorene (2-AAF) into N- and ring-hydroxylated metabolites and 2-aminofluorene at rates of 1.50, 9.73, and 1.98 nmol/mg cell protein/24 hr, respectively. Rats were i.v. injected with both LE-pEJ and LE-p17hGHneo carrying the hGH cDNA gene, and secreted hGH in the plasma which induced the synthesis of anti-hGH antibodies. A cell line was cloned from cultures of primary hepatocytes isolated from the liver of transfected rats. After 2 to 3 months in culture, this cell line secreted per day 18.9 micrograms albumin and 11.0 micrograms transferrin per 10(6) cells, 38.75 nmol total BA per mg cell protein, and up to 31 ng hGH per 10(6) cells without cloning hGH recombinant cells. A 24 hr control culture of primary hepatocytes isolated from non transfected rats secreted 25.5 micrograms albumin and 11.7 micrograms transferrin per 10(6) cells, and produced 21.64 nmol total BA and 2.13 nmol N-OH-2-AAF per mg cell protein. Hence, Ha-rasEJ transfection of either hepatocytes in vitro or liver cells in vivo, initiated cell cycles leading to presumptive proliferating hepatocytes which express liver function. |
| Databáze: | OpenAIRE |
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