An Optimized Method to Isolate Human Fibroblasts from Tissue for ex vivo Analysis

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Abstract

Despite their involvement in many physiological and pathological processes, fibroblasts remain a poorly-characterized cell type. Analysis of primary fibroblasts while maintaining their in vivo phenotype is challenging: standard methods for fibroblast isolation require cell culture in vitro, 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 ex vivo fibroblasts without the need for culture in vitro.

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Waise, S., Parker, R., Rose-Zerilli, M. J. J., Layfield, D. M., Wood, O., West, J., … Hanley, C. J. (2019). An Optimized Method to Isolate Human Fibroblasts from Tissue for ex vivo Analysis. Bio-Protocol, 9(23). https://doi.org/10.21769/BioProtoc.3440

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