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PMID: 3858822 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Maintenance of differentiated rat hepatocytes in primary culture.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 82 ·No. 10 ·1985-05-00 ·Pages 3252-6

Isom HC, Secott T, Georgoff I, Woodworth C, Mummaw J

Abstract

Normal adult rat hepatocytes remained viable and functional for at least 43 days when plated on collagen-coated dishes and fed chemically defined medium supplemented with dimethyl sulfoxide (Me2SO). Hepatocytes isolated by collagenase perfusion and cultured in the presence or absence of Me2SO were (i) examined by light and electron microscopy for morphological changes; (ii) analyzed for the production of albumin and other plasma proteins; and (iii) tested by autoradiography for DNA synthesis. Me2SO-treated cells continued to produce specific plasma proteins during the entire culture period; albumin production was consistently high (11-19 micrograms/ml of culture medium per 24 hr) from day 2 to at least day 43 after plating. Ultrastructural analyses demonstrated that Me2SO-treated hepatocytes resembled those from intact liver in organization of cytoplasmic organelles and cellular junctions. The optimal concentration for observing the morphological and biochemical effects of Me2SO was 2% (vol/vol). We conclude that supplementation of chemically defined medium with Me2SO enables maintenance of differentiated hepatocytes in culture for extended periods of time.

MeSH Terms
Albumins/metabolism Animals Cell Differentiation/drug effects Cells, Cultured Collagen Culture Media DNA/biosynthesis Dimethyl Sulfoxide/pharmacology Liver/cytology,metabolism Molecular Weight Proteins/metabolism Rats Tetradecanoylphorbol Acetate/pharmacology Time Factors
Chemicals
Albumins Culture Media Proteins Collagen DNA Tetradecanoylphorbol Acetate Dimethyl Sulfoxide
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Isom H C
Secott T
Georgoff I
Woodworth C
Mummaw J
References (35)
35 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.82.10.3252
Published
1985-05-00
Pages
3252-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC397753
Subset
IM
Grants
NCI NIH HHS · CA 09124 · United States
NCI NIH HHS · CA 18450 · United States
NCI NIH HHS · CA 23931 · United States
External Links
PubMed source DOI: 10.1073/pnas.82.10.3252 PMC Full Text View journal details
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