Profiling and authentication of human cell lines using short tandem repeat (STR) loci: Report from the National Cell Bank of Iran
Profiling and authentication of human cell lines using short tandem repeat (STR) loci: Report from the National Cell Bank of Iran
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DOI：
10.1016/j.biologicals.2006.10.001
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发表时间：
2007-06-01
期刊：
影响因子：
1.7
通讯作者：
Shokri, Fazel
中科院分区：
文献类型：
--
作者：
Azari, Shahram;Ahmadi, Nahid;Shokri, Fazel
确保细胞系的同质性和细胞污染检测能力是基于细胞的研究的最基本的步骤之一。短串联重复序列(STR)分析由于具有较高的鉴别效率、较低的成本和较高的可靠性，已成为人类细胞系鉴定的首选方法。在本研究中，利用13个基于联合DNA指数系统(CODIS)的STR和性别决定(釉质)基因建立了一种可重复性的方法，利用聚合酶链式反应(PCR)方法对保存在伊朗国家细胞库(NCBI)中的100个人类细胞系进行鉴定。聚合酶链式反应产物随后用聚丙烯酰胺凝胶电泳法(PAGE)分析，并通过银染显影，然后进行凝胶记录和软件分析。将获得的STR图谱与美国模式培养库(ATCC)和日本研究生物资源库(JCRB)的STR图谱进行比较，作为STR的参考。我们检测到NCBI人类细胞系之间有18.8%的交叉污染。据我们所知，这是首次使用13个CODIS核心短串联重复序列结合釉原蛋白鉴定人类细胞系。(C)2006年国际生物制品协会。爱思唯尔有限公司出版。保留所有权利。
Assurance of cell line homogeneity and capability of cell contamination detection are among the most essential steps of cell based research. Due to high discriminatory efficiency, low cost and reliability, analysis of short tandem repeats (STR) has been introduced as a method of choice for human cell line authentication. In the present study 13 Combined DNA Index System (CODIS) based STRs along with the gender determination (Amelogenin) gene were utilized to establish a reproducible approach for the authentication of 100 human cell lines deposited in the National Cell Bank of Iran (NCBI), using the polymerase chain reaction (PCR) method. PCR products were subsequently analyzed by polyacrylamide gel electrophoresis (PAGE) and visualized by silver staining followed by gel documentation and software analysis. STR profiles obtained were compared with those of the American Type Culture Collection (ATCC) and the Japanese Collection of Research Bioresource (JCRB) as STR references. We detected 18.8% cross contamination among the NCBI human cell lines. To our knowledge, this is the first report of authentication of human cell lines using the 13 CODIS core STRs combined with Amelogenin. (c) 2006 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.