In accordance with the 3 Rs principle (to replace, reduce and refine) animal models in biomedical research, we have developed and applied a new approach for sampling and analyzing hair follicles in various experimental settings. This involves use of a convenient device for non-invasive collection of hair follicles and processing methods that provide sufficient amounts of biological material to replace stressful and painful biopsies. Moreover, the main components of hair follicles are live cells of epithelial origin, which are highly relevant for most types of malignant tumors, so they provide opportunities for studying aging-related pathologies including cancer. Here, we report the successful use of the method to obtain mouse hair follicular cells for genotyping, quantitative PCR, and quantitative immunofluorescence. We present proof of concept data demonstrating its utility for routine genotyping and monitoring changes in quality and expression levels of selected proteins in mice after gamma irradiation and during natural or experimentally induced aging. We also performed pilot translation of animal experiments to human hair follicles irradiated ex vivo. Our results highlight the value of hair follicles as biological material for convenient in vivo sampling and processing in both translational research and routine applications, with a broad range of ethical and logistic advantages over currently used biopsy-based approaches.

Danish Cancer Society Research Center Copenhagen DK 2100 Denmark

Division of Genome Biology Department of Medical Biochemistry and Biophysics Science for Life Laboratory Karolinska Institute Stockholm 171 77 Sweden

Faculty of Science Palacky University and University Hospital in Olomouc Olomouc 779 00 Czech Republic

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University and University Hospital Olomouc Olomouc 779 00 Czech Republic

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Uncovering pre-cytokinetic block in cancer cells under shear stress using a disturbed flow-generating device

Cellular Senescence: Molecular Targets, Biomarkers, and Senolytic Drugs

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