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 language | English |
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Journal | In Vitro Cellular & Developmental Biology |
Volume | 24 |
Issue number | 11 |
Pages (from-to) | 1071-6 |
Number of pages | 6 |
ISSN | 0883-8364 |
DOIs | |
Publication status | Published - Nov 1988 |
Externally published | Yes |
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