Gene probes to detect cross-culture contamination in hormone producing cell lines.
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.
D007516:Adenoma, Islet Cell / Q000235:genetics*; D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D002871:Chromosome Banding; D006224:Cricetinae; D015342:DNA Probes; D006683:HLA-DQ Antigens / Q000235:genetics; D006801:Humans; D007328:Insulin / Q000235:genetics; D007340:Insulinoma / Q000235:genetics*; D008647:Mesocricetus; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D012091:Repetitive Sequences, Nucleic Acid
D015342:DNA Probes; D006683:HLA-DQ Antigens; D007328:Insulin