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Gene probes to detect cross-culture contamination in hormone producing cell lines

Matsuba, I. ; Lernmark, Å LU orcid ; Michelsen, B. ; Nielsen, J. H. LU ; Scholler, J. ; Vissing, H. ; Welinder, B. ; Tommerup, N. ; Mikkelsen, M. and Ishikawa, H. , et al. (1988) In In Vitro Cellular & Developmental Biology 24(11). p.1071-1076
Abstract

Cross-culture contamination of cell lines propagated in continuous culture is a frequent event and particularly difficult to resolve in cells expressing similar phenotypes. We demonstrate that DNA-DNA hybridization to blotted endonuclease-digested cell DNA effectively detects cross-culture contamination to monitor inter-species as well as intra-species cross contamination. An insulin-producing cell-line, Clone-16, originally cloned from a human fetal endocrine pancreatic cell line did not produce human c-peptide as anticipated. DNA from these cells showed no hybridization to the human ALU sequence probe, BLUR, and lacked restriction fragment length polymorphism typical for the human HLA-DQ β-chain gene. Although a human insulin gene... (More)

Cross-culture contamination of cell lines propagated in continuous culture is a frequent event and particularly difficult to resolve in cells expressing similar phenotypes. We demonstrate that DNA-DNA hybridization to blotted endonuclease-digested cell DNA effectively detects cross-culture contamination to monitor inter-species as well as intra-species cross contamination. An insulin-producing cell-line, Clone-16, originally cloned from a human fetal endocrine pancreatic cell line did not produce human c-peptide as anticipated. DNA from these cells showed no hybridization to the human ALU sequence probe, BLUR, and lacked restriction fragment length polymorphism typical for the human HLA-DQ β-chain gene. Although a human insulin gene probe showed a weak, nonhuman hybridization pattern, a cDNA probe for the Syrian hamster insulin gene hybridized strongly consistent with a single copy hamster insulin gene. Karyotyping confirmed the absence of human chromosomes in the Clone-16 cells while sizes, centromere indices, and banding patterns were identical to Syrian hamster fibroblasts. We conclude that the insulin-producing Clone-16 cells are of Syrian hamster origin and demonstrate the effective use of gene probes to control the origin of cell cultures.

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publishing date
type
Contribution to journal
publication status
published
keywords
ALU-probes, cross-culture contamination, DNA hybridization, insulin gene, insulinoma, karyotyping
in
In Vitro Cellular & Developmental Biology
volume
24
issue
11
pages
6 pages
publisher
Springer
external identifiers
  • scopus:0024269109
  • pmid:2903855
ISSN
0883-8364
DOI
10.1007/BF02620807
language
English
LU publication?
no
id
c50c431a-0ccd-416c-bd91-a7d6ed3ef305
date added to LUP
2019-09-11 10:02:15
date last changed
2024-03-13 08:09:18
@article{c50c431a-0ccd-416c-bd91-a7d6ed3ef305,
  abstract     = {{<p>Cross-culture contamination of cell lines propagated in continuous culture is a frequent event and particularly difficult to resolve in cells expressing similar phenotypes. We demonstrate that DNA-DNA hybridization to blotted endonuclease-digested cell DNA effectively detects cross-culture contamination to monitor inter-species as well as intra-species cross contamination. An insulin-producing cell-line, Clone-16, originally cloned from a human fetal endocrine pancreatic cell line did not produce human c-peptide as anticipated. DNA from these cells showed no hybridization to the human ALU sequence probe, BLUR, and lacked restriction fragment length polymorphism typical for the human HLA-DQ β-chain gene. Although a human insulin gene probe showed a weak, nonhuman hybridization pattern, a cDNA probe for the Syrian hamster insulin gene hybridized strongly consistent with a single copy hamster insulin gene. Karyotyping confirmed the absence of human chromosomes in the Clone-16 cells while sizes, centromere indices, and banding patterns were identical to Syrian hamster fibroblasts. We conclude that the insulin-producing Clone-16 cells are of Syrian hamster origin and demonstrate the effective use of gene probes to control the origin of cell cultures.</p>}},
  author       = {{Matsuba, I. and Lernmark, Å and Michelsen, B. and Nielsen, J. H. and Scholler, J. and Vissing, H. and Welinder, B. and Tommerup, N. and Mikkelsen, M. and Ishikawa, H. and Ikeda, Y. and Tanese, T.}},
  issn         = {{0883-8364}},
  keywords     = {{ALU-probes; cross-culture contamination; DNA hybridization; insulin gene; insulinoma; karyotyping}},
  language     = {{eng}},
  month        = {{11}},
  number       = {{11}},
  pages        = {{1071--1076}},
  publisher    = {{Springer}},
  series       = {{In Vitro Cellular & Developmental Biology}},
  title        = {{Gene probes to detect cross-culture contamination in hormone producing cell lines}},
  url          = {{http://dx.doi.org/10.1007/BF02620807}},
  doi          = {{10.1007/BF02620807}},
  volume       = {{24}},
  year         = {{1988}},
}