Gene probes to detect cross-culture contamination in hormone producing cell lines
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Gene probes to detect cross-culture contamination in hormone producing cell lines. / Matsuba, I; Lernmark, A; Madsen, Ole Dragsbæk; Michelsen, Birgitte; Nielsen, Jens Høiriis; Scholler, John; Vissing, H; Welinder, B; Tommerup, N; Mikkelsen, M.
I: In Vitro Cellular & Developmental Biology, Bind 24, Nr. 11, 11.1988, s. 1071-6.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › fagfællebedømt
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TY - JOUR
T1 - Gene probes to detect cross-culture contamination in hormone producing cell lines
AU - Matsuba, I
AU - Lernmark, A
AU - Madsen, Ole Dragsbæk
AU - Michelsen, Birgitte
AU - Nielsen, Jens Høiriis
AU - Scholler, John
AU - Vissing, H
AU - Welinder, B
AU - Tommerup, N
AU - Mikkelsen, M
PY - 1988/11
Y1 - 1988/11
N2 - 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.
AB - 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.
KW - Adenoma, Islet Cell
KW - Animals
KW - Blotting, Southern
KW - Cell Line
KW - Chromosome Banding
KW - Cricetinae
KW - DNA Probes
KW - HLA-DQ Antigens
KW - Humans
KW - Insulin
KW - Insulinoma
KW - Mesocricetus
KW - Polymorphism, Genetic
KW - Polymorphism, Restriction Fragment Length
KW - Repetitive Sequences, Nucleic Acid
U2 - 10.1007/BF02620807
DO - 10.1007/BF02620807
M3 - Journal article
C2 - 2903855
VL - 24
SP - 1071
EP - 1076
JO - In Vitro Cellular & Developmental Biology
JF - In Vitro Cellular & Developmental Biology
SN - 0883-8364
IS - 11
ER -
ID: 47974645