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保持清洁:国家转化医学科学中心的细胞培养质量控制经验。

Keeping It Clean: The Cell Culture Quality Control Experience at the National Center for Advancing Translational Sciences.

机构信息

National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, USA.

出版信息

SLAS Discov. 2020 Jun;25(5):491-497. doi: 10.1177/2472555220911451. Epub 2020 Apr 1.

Abstract

Quality control monitoring of cell lines utilized in biomedical research is of utmost importance and is critical for the reproducibility of data. Two key pitfalls in tissue culture are 1) cell line authenticity and 2) contamination. As a collaborative research institute, the National Center for Advancing Translational Sciences (NCATS) receives cell lines from a range of commercial and academic sources, which are adapted for high-throughput screening. Here, we describe the implementation of routine NCATS-wide testing and short tandem repeat (STR) testing for cell lines. Initial testing identified a >10% contamination rate. While the implementation of systematic testing has not fully suppressed contamination rates, clearly defined protocols that include the immediate destruction of contaminated cell lines wherever possible has enabled rapid intervention and removal of compromised cell lines. Data for >2000 cell line samples tested over 3 years, and case studies are provided. STR testing of 186 cell lines with established STR profiles revealed only five misidentified cell lines, all of which were received from external labs. The data collected over the 3 years since implementation of this systematic testing demonstrate the importance of continual vigilance for rapid identification of "problem" cell lines, for ensuring reproducible data in translational science research.

摘要

在生物医学研究中,细胞系的质量控制监测至关重要,这对于数据的可重复性至关重要。组织培养中有两个主要的陷阱:1)细胞系的真实性和 2)污染。作为一个合作研究机构,国家转化医学推进中心(NCATS)从各种商业和学术来源接收细胞系,这些细胞系经过了高通量筛选的适应。在这里,我们描述了常规的 NCATS 范围的细胞系测试和短串联重复(STR)测试的实施情况。初步测试确定了 >10%的污染率。虽然系统测试的实施并没有完全抑制污染率,但明确的协议,包括尽可能立即销毁污染的细胞系,已经能够实现快速干预和去除受损的细胞系。提供了超过 3 年的 >2000 个细胞系样本的测试数据和案例研究。对 186 个具有既定 STR 图谱的细胞系进行 STR 测试,只发现了 5 个鉴定错误的细胞系,这些细胞系均来自外部实验室。自实施这种系统测试以来的 3 年中收集的数据表明,持续保持警惕对于快速识别“问题”细胞系、确保转化科学研究中数据的可重复性非常重要。

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