Structural defects in DNA from primary hepatocyte cultures

Robyn D. Moir, Garry John Smith, Bernard W. Stewart

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Structural analysis of DNA from primary hepatocytes has been made by chromatography on benzoylated-DEAE-cellulose. DNA was labelled in vivo by incorporation of 3H-TdR after partial hepatectomy 2 weeks prior to animals being used to establish hepatocyte cultures. Structural change in DNA associated with the transition to primary culture was assessed by comparison of data with that obtained when DNA was isolated from the intact organ immediately after death. In terms of this comparison, DNA from primary hepatocytes was bound more extensively to benzoylated-DEAE-cellulose in the presence of 1.0 M NaCl. Thus 32% of primary hepatocyte DNA was recovered in the formamide-eluted fraction after chromatography, a result which reflected a higher proportion of DNA containing single-stranded regions. This proportion of formamide-eluted DNA in primary hepatocytes could not be further reduced by extensive shearing and could not be affected by manipulation of the isolation procedure. Analysis of DNA isolated from hepatocyte cell lines revealed structural characteristics similar to DNA isolated from intact liver. The data imply that extensive structural damage to DNA is a consequence of the primary status of the hepatocytes examined.

Original languageEnglish (US)
Pages (from-to)227-236
Number of pages10
JournalCell Biology International Reports
Volume7
Issue number3
DOIs
StatePublished - 1983
Externally publishedYes

Fingerprint

Hepatocytes
DNA
Chromatography
Single-Stranded DNA
Hepatectomy
DNA Damage
Cell Line
Liver

ASJC Scopus subject areas

  • Cell Biology

Cite this

Structural defects in DNA from primary hepatocyte cultures. / Moir, Robyn D.; Smith, Garry John; Stewart, Bernard W.

In: Cell Biology International Reports, Vol. 7, No. 3, 1983, p. 227-236.

Research output: Contribution to journalArticle

Moir, Robyn D. ; Smith, Garry John ; Stewart, Bernard W. / Structural defects in DNA from primary hepatocyte cultures. In: Cell Biology International Reports. 1983 ; Vol. 7, No. 3. pp. 227-236.
@article{e413df4a5aae4e9885cf5493e975473b,
title = "Structural defects in DNA from primary hepatocyte cultures",
abstract = "Structural analysis of DNA from primary hepatocytes has been made by chromatography on benzoylated-DEAE-cellulose. DNA was labelled in vivo by incorporation of 3H-TdR after partial hepatectomy 2 weeks prior to animals being used to establish hepatocyte cultures. Structural change in DNA associated with the transition to primary culture was assessed by comparison of data with that obtained when DNA was isolated from the intact organ immediately after death. In terms of this comparison, DNA from primary hepatocytes was bound more extensively to benzoylated-DEAE-cellulose in the presence of 1.0 M NaCl. Thus 32{\%} of primary hepatocyte DNA was recovered in the formamide-eluted fraction after chromatography, a result which reflected a higher proportion of DNA containing single-stranded regions. This proportion of formamide-eluted DNA in primary hepatocytes could not be further reduced by extensive shearing and could not be affected by manipulation of the isolation procedure. Analysis of DNA isolated from hepatocyte cell lines revealed structural characteristics similar to DNA isolated from intact liver. The data imply that extensive structural damage to DNA is a consequence of the primary status of the hepatocytes examined.",
author = "Moir, {Robyn D.} and Smith, {Garry John} and Stewart, {Bernard W.}",
year = "1983",
doi = "10.1016/0309-1651(83)90230-8",
language = "English (US)",
volume = "7",
pages = "227--236",
journal = "Cell Biology International",
issn = "1065-6995",
publisher = "Portland Press Ltd.",
number = "3",

}

TY - JOUR

T1 - Structural defects in DNA from primary hepatocyte cultures

AU - Moir, Robyn D.

AU - Smith, Garry John

AU - Stewart, Bernard W.

PY - 1983

Y1 - 1983

N2 - Structural analysis of DNA from primary hepatocytes has been made by chromatography on benzoylated-DEAE-cellulose. DNA was labelled in vivo by incorporation of 3H-TdR after partial hepatectomy 2 weeks prior to animals being used to establish hepatocyte cultures. Structural change in DNA associated with the transition to primary culture was assessed by comparison of data with that obtained when DNA was isolated from the intact organ immediately after death. In terms of this comparison, DNA from primary hepatocytes was bound more extensively to benzoylated-DEAE-cellulose in the presence of 1.0 M NaCl. Thus 32% of primary hepatocyte DNA was recovered in the formamide-eluted fraction after chromatography, a result which reflected a higher proportion of DNA containing single-stranded regions. This proportion of formamide-eluted DNA in primary hepatocytes could not be further reduced by extensive shearing and could not be affected by manipulation of the isolation procedure. Analysis of DNA isolated from hepatocyte cell lines revealed structural characteristics similar to DNA isolated from intact liver. The data imply that extensive structural damage to DNA is a consequence of the primary status of the hepatocytes examined.

AB - Structural analysis of DNA from primary hepatocytes has been made by chromatography on benzoylated-DEAE-cellulose. DNA was labelled in vivo by incorporation of 3H-TdR after partial hepatectomy 2 weeks prior to animals being used to establish hepatocyte cultures. Structural change in DNA associated with the transition to primary culture was assessed by comparison of data with that obtained when DNA was isolated from the intact organ immediately after death. In terms of this comparison, DNA from primary hepatocytes was bound more extensively to benzoylated-DEAE-cellulose in the presence of 1.0 M NaCl. Thus 32% of primary hepatocyte DNA was recovered in the formamide-eluted fraction after chromatography, a result which reflected a higher proportion of DNA containing single-stranded regions. This proportion of formamide-eluted DNA in primary hepatocytes could not be further reduced by extensive shearing and could not be affected by manipulation of the isolation procedure. Analysis of DNA isolated from hepatocyte cell lines revealed structural characteristics similar to DNA isolated from intact liver. The data imply that extensive structural damage to DNA is a consequence of the primary status of the hepatocytes examined.

UR - http://www.scopus.com/inward/record.url?scp=0020732031&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020732031&partnerID=8YFLogxK

U2 - 10.1016/0309-1651(83)90230-8

DO - 10.1016/0309-1651(83)90230-8

M3 - Article

C2 - 6850855

AN - SCOPUS:0020732031

VL - 7

SP - 227

EP - 236

JO - Cell Biology International

JF - Cell Biology International

SN - 1065-6995

IS - 3

ER -