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Authentication of primary murine cell lines by a microfluidics-based lab-on-chip system

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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. and Singh, N. and Bamopoulos, S. and Gjerga, E. and Schmalbrock, L.K. and Balabanian, K. and Schick, M. and 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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