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

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

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.
OriginalsprogEngelsk
TidsskriftIn vitro Cellular and Developmental Biology
Vol/bind24
Udgave nummer11
Sider (fra-til)1071-6
Antal sider6
StatusUdgivet - nov. 1988

ID: 47974645