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

I Matsuba, A Lernmark, Ole Dragsbæk Madsen, Birgitte Michelsen, Jens Høiriis Nielsen, John Scholler, H Vissing, B Welinder, N Tommerup, M Mikkelsen

Research output: Contribution to journalJournal articleResearchpeer-review

1 Citation (Scopus)

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 beta-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.
Original languageEnglish
JournalIn Vitro Cellular & Developmental Biology
Volume24
Issue number11
Pages (from-to)1071-6
Number of pages6
ISSN0883-8364
DOIs
Publication statusPublished - Nov 1988
Externally publishedYes

Keywords

  • Adenoma, Islet Cell
  • Animals
  • Blotting, Southern
  • Cell Line
  • Chromosome Banding
  • Cricetinae
  • DNA Probes
  • HLA-DQ Antigens
  • Humans
  • Insulin
  • Insulinoma
  • Mesocricetus
  • Polymorphism, Genetic
  • Polymorphism, Restriction Fragment Length
  • Repetitive Sequences, Nucleic Acid

Cite this