Matsuba I, Lernmark A, Madsen O, Michelsen B, Nielsen J H, Scholler J, Vissing H, Welinder B, Tommerup N, Mikkelsen M
Hagedorn Research Laboratory, Gentofte, Denmark.
In Vitro Cell Dev Biol. 1988 Nov;24(11):1071-6. doi: 10.1007/BF02620807.
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.
在连续培养中传代的细胞系发生跨文化污染是常有的事，在表达相似表型的细胞中尤其难以解决。我们证明，与印迹的经核酸内切酶消化的细胞DNA进行DNA-DNA杂交能有效检测跨文化污染，以监测种间以及种内交叉污染。一个最初从人胎儿内分泌胰腺细胞系克隆而来的产胰岛素细胞系Clone-16，并未如预期那样产生人C肽。这些细胞的DNA与人类ALU序列探针BLUR没有杂交信号，并且缺乏人类HLA-DQβ链基因典型的限制性片段长度多态性。尽管一个人胰岛素基因探针显示出微弱的非人类杂交模式，但一个叙利亚仓鼠胰岛素基因的cDNA探针与之强烈杂交，这与单拷贝仓鼠胰岛素基因一致。核型分析证实Clone-16细胞中不存在人类染色体，而其大小、着丝粒指数和带型与叙利亚仓鼠成纤维细胞相同。我们得出结论，产胰岛素的Clone-16细胞源自叙利亚仓鼠，并证明了基因探针在控制细胞培养物来源方面的有效应用。
