SBIR-STTR Award

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.

Public Health Relevance Statement:
Narrative The goal of the project is to develop new kits for detecting low levels of cross-contaminations in human cell cultures and prevent contamination from leading to irreproducible research results or incorrect scientific conclu- sions. Current technologies are often labor intensive, slow, costly, and/or not suited for cancer cell lines. Building upon NuProbe’s proprietary technology, we propose to develop rapid, cost effective, reliable, and easy-to-use cell line contamination detection kits that will be compatible with commonly used laboratory instruments.

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

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.

Public Health Relevance Statement:
The goal of the project is to develop new kits for detecting low levels of cross-contaminations in human cell cultures and prevent contamination from leading to irreproducible research results or incorrect scientific conclusions. Current technologies are often labor intensive, slow, costly, and/or not suited for cancer cell lines. Building upon NuProbe’s proprietary technology, we propose to develop rapid, cost effective, reliable, and easy-to-use cell line contamination detection kits that will be compatible with commonly used laboratory instruments.

NIH Spending Category:
Aging; Biotechnology; Genetics

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