1. Authentication of Primary Murine Cell Lines by a Microfluidics-Based Lab-On-Chip System
- Author
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Hong, Yingfen, Singh, Nikita, Bamopoulos, Stefanos, Gjerga, Enio, Schmalbrock, Laura K., Balabanian, Karl, Schick, Markus, Keller, Ulrich, and Wirth, Matthias
- Subjects
Cancer Research ,lcsh:Biology (General) ,FLA ,HoxB8 ,authentication ,mouse cell lines ,bioanalyzer ,lcsh:QH301-705.5 ,STR-PCR ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit ,Article - Abstract
The reliable authentication of cell lines is a prerequisite for the reproducibility and replicability of experiments. A common method of cell line authentication is the fragment length analysis (FLA) of short-tandem repeats (STR) by capillary electrophoresis. However, this technique is not always accessible and is often costly. Using a microfluidic electrophoresis system, we analyzed the quality and integrity of different murine cell lines by STR profiling. As a proof of concept, we isolated and immortalized hematopoietic progenitor cells (HPC) of various genotypes through retroviral transduction of the fusion of the estrogen receptor hormone-binding domain with the coding sequence of HoxB8. Cell lines were maintained in the HPC state with Flt3 ligand (FL) and estrogen treatment and could be characterized upon differentiation. In a validation cohort, we applied this technique on primary mutant Kras-driven pancreatic cancer cell lines, which again allowed for clear discrimination. In summary, our study provides evidence that FLA of STR-amplicons by microfluidic electrophoresis allows for stringent quality control and the tracking of cross-contaminations in both genetically stable HPC lines and cancer cell lines, making it a simple and cost-efficient alternative to traditional capillary electrophoresis.
- Published
- 2020