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Rapid, Simple and Cost-Effective Detection of Human Cell Line Contamination

快速、简单且经济高效地检测人类细胞系污染

基本信息

批准号：
10004552
负责人：
Yan Helen Yan
金额：
$ 25.5万
依托单位：
NUPROBE USA, INC.
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-30 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10004552
关键词：
Affect Aneuploidy Awareness Benchmarking Biological Assay Biomedical Research Budgets Cancer cell line Cancerous Capillary Electrophoresis Caring Cell Culture Techniques Cell Line Cell physiology Cells Communities Computer software Consumption Detection Development Disease Ensure Environmental Risk Factor Equipment Exhibits Gene Frequency Genomic Instability Genotype Goals Hela Cells Hour Human Human Cell Line Immunoglobulin Variable Region Knowledge Laboratories Length Loss of Heterozygosity Measures Methods Microsatellite Instability Microsatellite Repeats Minor Modeling Monitor Performance Phase Polymerase Chain Reaction Protocols documentation Reaction Reference Standards Reproducibility Research Research Contracts Rice Sampling Science Series Short Tandem Repeat Side Single Nucleotide Polymorphism Site Standardization Statistical Data Interpretation Stress Tests Technology Testing Time Translating Universities Validation base cancer cell cost cost effective design drug candidate experimental study instrument laboratory equipment prevent rare variant research in practice service providers virtual

项目摘要

Cell line contamination and misidentification have dramatic repercussions in the field biomedical research, largely contributing to the growing concerns about irreproducible results and posing a significant burden on the global biomedical research budget. While the general awareness about these issues has raised over the last years, current authentication technologies such as short tandem repeat (STR) profiling are often labor intensive, slow, costly and not suited for cancer cell lines, which make it very difficult to reliably and continuously monitor human cell cultures. Profiling single nucleotide polymorphisms (SNPs) would provide a good alternative to STR for monitoring cancer cell cultures, SNPs being less affected by genomic instabilities than STR markers. NuProbe’s blocker displacement amplification (BDA) technology uniquely enables multiplexed rare allele enrichment by PCR. Building upon NuProbe’s BDA technology, we propose to develop a series of highly multiplexed and highly sensitive qPCR-based SNP profiling assays to detect intra-species cross-contaminants present in human cell cultures at low levels (down to 1%). We anticipate these assays to translate into rapid, cost effective, reliable, and easy-to-use cell line contamination detection kits compatible with commonly available qPCR instruments.

细胞系污染和错误鉴定在生物医学研究领域具有巨大的影响，这在很大程度上加剧了人们对不可复制的结果的日益担忧，并对全球生物医学研究预算。虽然在过去的一段时间里，人们对这些问题的普遍认识有所提高，多年来，目前的鉴定技术如短串联重复序列（STR）分析通常是劳动密集型的，缓慢、昂贵且不适用于癌细胞系，这使得非常难以可靠和连续地监测人类细胞培养物。单核苷酸多态性（SNPs）分析将提供一个很好的替代STR 用于监测癌细胞培养物，SNP比STR标记受基因组不稳定性的影响更小。 NuProbe的阻断剂置换扩增（BDA）技术独特地实现了多重稀有等位基因通过PCR富集。基于NuProbe的BDA技术，我们建议开发一系列高度多重和高灵敏度的基于qPCR的SNP分析测定，用于检测种内交叉污染物在人类细胞培养中以低水平存在（低至1%）。我们预计这些检测将转化为快速，具有成本效益、可靠且易于使用的细胞系污染检测试剂盒， qPCR仪器。