Chronobiology and Sleep Research Group, Institute of Pharmacology and Toxicology, University of Zurich, Zurich, Switzerland.
Methods Mol Biol. 2021;2130:53-67. doi: 10.1007/978-1-0716-0381-9_4.
Human cells, especially primary fibroblasts from skin punch biopsy, have emerged over the last decade as powerful, unlimited, and easily accessible resources that bridge the gap between animal models and human subjects in basic as well as clinical research. The cells also retain molecular circadian clocks that reflect subject-specific differences in circadian physiology, and the cellular rhythms can be measured easily in large scale. This is a series of protocols that describes the procedure to measure circadian rhythms in these cells, starting from deriving fibroblasts from skin punch biopsy, to generation of stable cells expressing a circadian reporter, and finally measurement of cellular rhythms in large scale.
在过去的十年中,人类细胞,尤其是从皮肤穿刺活检中提取的原代成纤维细胞,已经成为强大的、无限的、易于获取的资源,在基础研究和临床研究中弥合了动物模型和人类受试者之间的差距。这些细胞还保留了分子生物钟,反映了生物钟生理学中的个体差异,并且可以很容易地在大规模水平上测量细胞节律。这是一系列描述从皮肤穿刺活检中提取成纤维细胞,生成表达生物钟报告基因的稳定细胞,最后在大规模水平上测量细胞节律的方案。