Waise Sara, Parker Rachel, Rose-Zerilli Matthew J J, Layfield David M, Wood Oliver, West Jonathan, Ottensmeier Christian H, Thomas Gareth J, Hanley Christopher J
Cancer Sciences Unit, University of Southampton, UK.
Institute for Life Sciences, University of Southampton, UK.
Bio Protoc. 2019 Dec 5;9(23):e3440. doi: 10.21769/BioProtoc.3440.
Despite their involvement in many physiological and pathological processes, fibroblasts remain a poorly-characterized cell type. Analysis of primary fibroblasts while maintaining their phenotype is challenging: standard methods for fibroblast isolation require cell culture , which is known to alter phenotypes. Previously-described protocols for the dissociation of primary tissues fail to extract sufficient numbers of fibroblasts, instead largely yielding immune cells. Here, we describe an optimized method for generating a fibroblast-enriched single-cell suspension from human tissues using combined mechanical and enzymatic dissociation. This allows analysis of fibroblasts without the need for culture .
尽管成纤维细胞参与了许多生理和病理过程，但它们仍然是一种特征尚不明确的细胞类型。在维持原代表型的同时对原代成纤维细胞进行分析具有挑战性：成纤维细胞分离的标准方法需要细胞培养，而众所周知，细胞培养会改变细胞表型。先前描述的原代组织解离方案无法提取足够数量的成纤维细胞，反而主要产生免疫细胞。在这里，我们描述了一种优化的方法，使用机械和酶解相结合的方式从人类组织中生成富含成纤维细胞的单细胞悬液。这使得无需培养即可对成纤维细胞进行分析。
