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Title: Regulated expression of erythropoietin by two human hepatoma cell lines

Abstract

The development of a cell culture system that produces erythropoietin (Epo) in a regulated manner has been the focus of much effort. The authors have screened multiple renal and hepatic cell lines for either constitutive or regulated expression of Epo. Only the human hepatoma cell lines, Hep3B and HepG2, made significant amounts of Epo as measured both by radioimmunoassay and in vitro bioassay (as much as 330 milliunits per 10/sup 6/ cells in 24 hr). The constitutive production of Epo increased dramatically as a function of cell density in both cell lines. At cell densities < 3.3 x 10/sup 5/ cells per cm/sup 2/, there was little constitutive release of Epo in the medium. With Hep3B cells grown at low cell densities, a mean 18-fold increase in Epo expression was seen in response to hypoxia and a 6-fold increase was observed in response to incubation in medium containing 50 ..mu..M cobalt(II) chloride. At similar low cell densities, Epo production in HepG2 cells could be enhanced an average of about 3-fold by stimulation with either hypoxia or cobalt(II) chloride. Upon such stimulation, both cell lines demonstrated markedly elevated levels of Epo mRNA. Hence, both Hep3B and HepG2 cell lines provide anmore » excellent in vitro system in which to study the physiological regulation of Epo expression.« less

Authors:
; ; ;
Publication Date:
Research Org.:
Howard Hughes Medical Institute, Boston, MA (USA)
OSTI Identifier:
6879466
Resource Type:
Journal Article
Journal Name:
Proc. Natl. Acad. Sci. U.S.A.; (United States)
Additional Journal Information:
Journal Volume: 84:22
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ERYTHROPOIETIN; BIOCHEMISTRY; RADIOIMMUNOASSAY; CELL CULTURES; CYTOLOGICAL TECHNIQUES; HEPATOMAS; HYBRIDIZATION; IODINE 125; PHOSPHORUS 32; RNA; TUMOR CELLS; ANIMAL CELLS; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; CHEMISTRY; DAYS LIVING RADIOISOTOPES; DISEASES; ELECTRON CAPTURE RADIOISOTOPES; IMMUNOASSAY; IMMUNOLOGY; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; LIGHT NUCLEI; MITOGENS; NEOPLASMS; NUCLEI; NUCLEIC ACIDS; ODD-EVEN NUCLEI; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; PHOSPHORUS ISOTOPES; PROTEINS; RADIOASSAY; RADIOIMMUNOLOGY; RADIOISOTOPES; TRACER TECHNIQUES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Goldberg, M A, Glass, G A, Cunningham, J M, and Bunn, H F. Regulated expression of erythropoietin by two human hepatoma cell lines. United States: N. p., 1987. Web. doi:10.1073/pnas.84.22.7972.
Goldberg, M A, Glass, G A, Cunningham, J M, & Bunn, H F. Regulated expression of erythropoietin by two human hepatoma cell lines. United States. https://doi.org/10.1073/pnas.84.22.7972
Goldberg, M A, Glass, G A, Cunningham, J M, and Bunn, H F. 1987. "Regulated expression of erythropoietin by two human hepatoma cell lines". United States. https://doi.org/10.1073/pnas.84.22.7972.
@article{osti_6879466,
title = {Regulated expression of erythropoietin by two human hepatoma cell lines},
author = {Goldberg, M A and Glass, G A and Cunningham, J M and Bunn, H F},
abstractNote = {The development of a cell culture system that produces erythropoietin (Epo) in a regulated manner has been the focus of much effort. The authors have screened multiple renal and hepatic cell lines for either constitutive or regulated expression of Epo. Only the human hepatoma cell lines, Hep3B and HepG2, made significant amounts of Epo as measured both by radioimmunoassay and in vitro bioassay (as much as 330 milliunits per 10/sup 6/ cells in 24 hr). The constitutive production of Epo increased dramatically as a function of cell density in both cell lines. At cell densities < 3.3 x 10/sup 5/ cells per cm/sup 2/, there was little constitutive release of Epo in the medium. With Hep3B cells grown at low cell densities, a mean 18-fold increase in Epo expression was seen in response to hypoxia and a 6-fold increase was observed in response to incubation in medium containing 50 ..mu..M cobalt(II) chloride. At similar low cell densities, Epo production in HepG2 cells could be enhanced an average of about 3-fold by stimulation with either hypoxia or cobalt(II) chloride. Upon such stimulation, both cell lines demonstrated markedly elevated levels of Epo mRNA. Hence, both Hep3B and HepG2 cell lines provide an excellent in vitro system in which to study the physiological regulation of Epo expression.},
doi = {10.1073/pnas.84.22.7972},
url = {https://www.osti.gov/biblio/6879466}, journal = {Proc. Natl. Acad. Sci. U.S.A.; (United States)},
number = ,
volume = 84:22,
place = {United States},
year = {Sun Nov 01 00:00:00 EST 1987},
month = {Sun Nov 01 00:00:00 EST 1987}
}