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

Research output: Contribution to journalArticle

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 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.

Details

Authors
  • I. Matsuba
  • Å Lernmark
  • B. Michelsen
  • J. H. Nielsen
  • J. Scholler
  • H. Vissing
  • B. Welinder
  • N. Tommerup
  • M. Mikkelsen
  • H. Ishikawa
  • Y. Ikeda
  • T. Tanese
External organisations
  • Hagedorn Research Institute
  • Kennedy Institute - National Eye Clinic
  • Jikei University School of Medicine
Research areas and keywords

Keywords

  • ALU-probes, cross-culture contamination, DNA hybridization, insulin gene, insulinoma, karyotyping
Original languageEnglish
Pages (from-to)1071-1076
Number of pages6
JournalIn Vitro Cellular & Developmental Biology
Volume24
Issue number11
Publication statusPublished - 1988 Nov 1
Publication categoryResearch
Peer-reviewedYes
Externally publishedYes