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Castello-Cros R , Cukierman E
Stromagenesis during tumorigenesis: characterization of tumor-associated fibroblasts and stroma-derived 3D matrices
Methods Mol Biol. 2009 ;522 :275-305
PMID: 19247611    PMCID: PMC2670062   
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It is increasingly recognized that interactions between cancer cells and their surrounding stroma are critical for promoting the growth and invasiveness of tumors. For example, cancer cells alter the topography and molecular composition of stromal extracellular matrix by increasing paracrine regulation of fibroblastic stromal cells during early tumor development. In turn, these physical and biochemical alterations of the stroma, profoundly affect the properties of the cancer cells. However, little is known about the cross-talk between stroma and cancer cells, and it is mainly due to the lack of a suitable in vitro system to mimic the stroma in vivo. We present an in vivo-like 3D stromal system derived from fibroblasts harvested from tissue samples representing various stages of stroma progression during tumorigenesis. The chapter describes how to isolate and characterize fibroblasts from a plethora of tissue samples. It describes how to produce and characterize fibroblast-derived 3D matrices. Finally, it describes how to test matrix permissiveness by analyzing the morphology of cancer cells cultured within various 3D matrices.
Castello-Cros, Remedios Cukierman, Edna CA006927/CA/NCI NIH HHS/United States R01-CA113451/CA/NCI NIH HHS/United States Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't United States Methods in molecular biology (Clifton, N.J.) Methods Mol Biol. 2009;522:275-305.