Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University and University Hospital Olomouc, Olomouc 779 00, Czech Republic.
Faculty of Science, Palacky University and University Hospital in Olomouc, Olomouc 779 00, Czech Republic.
Aging (Albany NY). 2021 Dec 6;13(23):25004-25024. doi: 10.18632/aging.203744.
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 . Our results highlight the value of hair follicles as biological material for convenient 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.
根据 3R 原则（替代、减少和优化），我们在生物医学研究中开发并应用了一种新方法，用于在各种实验环境中采样和分析毛囊。这涉及使用一种方便的设备进行非侵入性的毛囊采集，以及提供足够生物材料的处理方法，以替代有压力和痛苦的活检。此外，毛囊的主要成分是上皮来源的活细胞，这些细胞与大多数类型的恶性肿瘤高度相关，因此为研究包括癌症在内的与衰老相关的病理提供了机会。在这里，我们报告了成功使用该方法从毛囊中获得小鼠毛囊细胞进行基因分型、定量 PCR 和定量免疫荧光的结果。我们提供了概念验证数据，证明了该方法在常规基因分型以及监测γ辐射后和自然或实验性衰老过程中，小鼠中选定蛋白质的质量和表达水平变化方面的实用性。我们还对动物实验进行了初步的翻译，将其应用于人类毛囊的辐射实验。我们的结果突出了毛囊作为生物材料的价值，它便于取材和处理，与目前基于活检的方法相比，具有广泛的伦理和后勤优势。
