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聚合酶链反应鉴定和验证啮齿动物细胞系。

Identification and verification of rodent cell lines by polymerase chain reaction.

机构信息

Department of Human and Animal Cell Cultures, DSMZ-German Collection of Microorganisms and Cell Cultures, Inhoffenstrasse 7B, 38124, Braunschweig, Germany,

出版信息

Cytotechnology. 2008 Jan;56(1):49-56. doi: 10.1007/s10616-007-9106-z. Epub 2007 Oct 31.

Abstract

Cell lines represent valuable tools for basic research and diagnostic applications as well as for the production of biological products such as antibodies or vaccines. For all cell culturists, a well-identified origin of their cell lines as well as the periodic re-examination of their identity should be a basic requirement. We established a simple polymerase chain reaction (PCR) to verify or identify rodent and human cell lines. Since mouse-, rat-, Chinese hamster- and Syrian hamster-derived cell lines represent the most frequently used rodent cell lines, our investigations were focused on these species. Our assay used oligonucleotide primers annealing to sequences within the beta-actin and the beta-globin gene and to repetitive DNA. Primers were designed mostly from intron sequences of the genes aiming to amplify only one specific DNA segment and thus enabling to exclude easily false DNA. More than 130 cells lines originating from the five species were analyzed in that study. Our PCR revealed specific profiles for all species investigated. No further methods like DNA sequencing or fragment length polymorphism analysis were needed to differentiate these species. The results introduce our PCR-assay as a rapid, specific and routinely feasible tool in order to identify or distinguish rodent cell lines from each other and from human cell lines.

摘要

细胞系是基础研究和诊断应用以及生产抗体或疫苗等生物产品的有价值的工具。对于所有细胞培养者来说,其细胞系的来源明确以及定期对其身份进行重新检查应该是基本要求。我们建立了一种简单的聚合酶链反应(PCR)来验证或鉴定啮齿动物和人类细胞系。由于小鼠、大鼠、中国仓鼠和叙利亚仓鼠来源的细胞系是最常用的啮齿动物细胞系,因此我们的研究集中在这些物种上。我们的检测使用寡核苷酸引物与β-肌动蛋白和β-球蛋白基因内的序列以及重复 DNA 退火。引物主要设计自基因的内含子序列,旨在扩增仅一个特定的 DNA 片段,从而可以轻松排除假 DNA。在该研究中分析了来自五个物种的 130 多个细胞系。我们的 PCR 揭示了所有被研究物种的特定图谱。无需进一步的方法,如 DNA 测序或片段长度多态性分析,即可区分这些物种。这些结果表明,我们的 PCR 检测方法是一种快速、特异性和常规可行的工具,可用于识别或区分彼此以及与人类细胞系的啮齿动物细胞系。

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本文引用的文献

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