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| Item Type: | Article |
|---|---|
| Title: | Authentication of primary murine cell lines by a microfluidics-based lab-on-chip system |
| Creators Name: | Hong, Y., Singh, N., Bamopoulos, S., Gjerga, E., Schmalbrock, L.K., Balabanian, K., Schick, M., Keller, U. and Wirth, M. |
| 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. |
| Keywords: | Authentication, FLA, STR-PCR, Bioanalyzer, Mouse Cell Lines, HoxB8, Animals, Mice |
| Source: | Biomedicines |
| ISSN: | 2227-9059 |
| Publisher: | MDPI |
| Volume: | 8 |
| Number: | 12 |
| Page Range: | 590 |
| Date: | 9 December 2020 |
| Official Publication: | https://doi.org/10.3390/biomedicines8120590 |
| PubMed: | View item in PubMed |
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