Cancer-associated fibroblasts (CAFs) represent a crucial component of cancer microenvironment. CAFs significantly influence biological properties of various types of cancer in terms of local aggressiveness, recurrence, and metastatic behaviour. This chapter summarizes a simple protocol for isolation of normal fibroblasts and their cancer-associated counterparts from normal human skin and mucosa, respectively, as well as from samples of human tumours such as basal/squamous carcinoma, melanoma, and breast cancer, and employment of this procedure for other types of cancer is possible. Isolated fibroblasts can be expanded in vitro and employed for further analysis of, e.g., DNA, RNA, protein, etc.
Cancer microenvironment Cancer-associated fibroblast Cell cultivation Fibroblast Fibroblast isolation
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This chapter was supported by the Grant Agency of the Czech Republic (grant no. 16-05534S); Charles University (project of PROGRES Q28/F1); Ministry of Education, Youth, and Sports of CR within the National Sustainability Program II (project BIOCEV-FAR; registration no. LQ1604); and project BIOCEV (grant no. CZ.1.05/1.1.00/02.0109). The present study is a result of the following project implementation: the equipment for metabolomic and cell analyses (grant no. CZ.1.05/2.1.00/19.0400) supported by the Research and Development for Innovations Operational Program, co-financed by the European regional development fund and the state budget of the Czech Republic.
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