Inhibitor of protein degradation formed during incubation of isolated rat hepatocytes in a cell culture medium. Its identification as ammonia

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Isolated rat hepatocytes, incubated as a suspension in a tissue culture medium (Dulbecco's), are in negative nitrogen balance and exhibit a continuous net release of amino acids and/or urea. The cells also generate ammonia and/or urea by the deamination of medium glutamine. Under normoxic conditions, ammonia is rapidly converted to urea, but under hypoxic conditions, arising, e.g., if the cells are allowed to sediment, ammonia accumulates. By interference with the analytical procedures and the calculation of nitrogen balance, the accumulation of ammonia may give a false impression of cellular hyper-catabolic activity during the first hour of incubation. When this effect has subsided (or been corrected for), it becomes evident that the accumulated ammonia exerts an inhibitory influence on protein degradation, reducing both the total nitrogen release and the liberation of [14C]valine from pre-labelled liver cell protein. The fact that protein degradation is inhibited by ammonia may be of considerable importance for the growth of cells under tissue culture conditions, where ammonia is so readily generated by the spontaneous or metabolic deamination of glutamine.

Original languageEnglish (US)
Pages (from-to)207-217
Number of pages11
JournalExperimental Cell Research
Volume107
Issue number1
DOIs
StatePublished - Jan 1 1977

Fingerprint

Ammonia
Proteolysis
Culture Media
Hepatocytes
Cell Culture Techniques
Urea
Deamination
Nitrogen
Glutamine
Valine
Suspensions
Amino Acids
Liver
Growth
Proteins

ASJC Scopus subject areas

  • Cell Biology

Cite this

@article{c419200d93ae4026acaf76c31124431b,
title = "Inhibitor of protein degradation formed during incubation of isolated rat hepatocytes in a cell culture medium. Its identification as ammonia",
abstract = "Isolated rat hepatocytes, incubated as a suspension in a tissue culture medium (Dulbecco's), are in negative nitrogen balance and exhibit a continuous net release of amino acids and/or urea. The cells also generate ammonia and/or urea by the deamination of medium glutamine. Under normoxic conditions, ammonia is rapidly converted to urea, but under hypoxic conditions, arising, e.g., if the cells are allowed to sediment, ammonia accumulates. By interference with the analytical procedures and the calculation of nitrogen balance, the accumulation of ammonia may give a false impression of cellular hyper-catabolic activity during the first hour of incubation. When this effect has subsided (or been corrected for), it becomes evident that the accumulated ammonia exerts an inhibitory influence on protein degradation, reducing both the total nitrogen release and the liberation of [14C]valine from pre-labelled liver cell protein. The fact that protein degradation is inhibited by ammonia may be of considerable importance for the growth of cells under tissue culture conditions, where ammonia is so readily generated by the spontaneous or metabolic deamination of glutamine.",
author = "Seglen, {Per Ottar}",
year = "1977",
month = "1",
day = "1",
doi = "10.1016/0014-4827(77)90402-5",
language = "English (US)",
volume = "107",
pages = "207--217",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Inhibitor of protein degradation formed during incubation of isolated rat hepatocytes in a cell culture medium. Its identification as ammonia

AU - Seglen, Per Ottar

PY - 1977/1/1

Y1 - 1977/1/1

N2 - Isolated rat hepatocytes, incubated as a suspension in a tissue culture medium (Dulbecco's), are in negative nitrogen balance and exhibit a continuous net release of amino acids and/or urea. The cells also generate ammonia and/or urea by the deamination of medium glutamine. Under normoxic conditions, ammonia is rapidly converted to urea, but under hypoxic conditions, arising, e.g., if the cells are allowed to sediment, ammonia accumulates. By interference with the analytical procedures and the calculation of nitrogen balance, the accumulation of ammonia may give a false impression of cellular hyper-catabolic activity during the first hour of incubation. When this effect has subsided (or been corrected for), it becomes evident that the accumulated ammonia exerts an inhibitory influence on protein degradation, reducing both the total nitrogen release and the liberation of [14C]valine from pre-labelled liver cell protein. The fact that protein degradation is inhibited by ammonia may be of considerable importance for the growth of cells under tissue culture conditions, where ammonia is so readily generated by the spontaneous or metabolic deamination of glutamine.

AB - Isolated rat hepatocytes, incubated as a suspension in a tissue culture medium (Dulbecco's), are in negative nitrogen balance and exhibit a continuous net release of amino acids and/or urea. The cells also generate ammonia and/or urea by the deamination of medium glutamine. Under normoxic conditions, ammonia is rapidly converted to urea, but under hypoxic conditions, arising, e.g., if the cells are allowed to sediment, ammonia accumulates. By interference with the analytical procedures and the calculation of nitrogen balance, the accumulation of ammonia may give a false impression of cellular hyper-catabolic activity during the first hour of incubation. When this effect has subsided (or been corrected for), it becomes evident that the accumulated ammonia exerts an inhibitory influence on protein degradation, reducing both the total nitrogen release and the liberation of [14C]valine from pre-labelled liver cell protein. The fact that protein degradation is inhibited by ammonia may be of considerable importance for the growth of cells under tissue culture conditions, where ammonia is so readily generated by the spontaneous or metabolic deamination of glutamine.

UR - http://www.scopus.com/inward/record.url?scp=0017738395&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017738395&partnerID=8YFLogxK

U2 - 10.1016/0014-4827(77)90402-5

DO - 10.1016/0014-4827(77)90402-5

M3 - Article

C2 - 16760

AN - SCOPUS:0017738395

VL - 107

SP - 207

EP - 217

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 1

ER -