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Analysis of Cancer Stromal Reaction Using an O-ring Co-culture Assay.

Bio-protocol (2017-02-20)
Vivien Jane Coulson-Thomas
ABSTRACT

We have developed a 2D heterotypic co-culture technique between fibroblasts and cancer cells that enables the study of the stromal reaction. For such, stromal cells are seeded and cultured immediately around a tumour cell line, and the cells establish cell-cell contacts, as well as a gradient of soluble factors throughout the stromal cells, similar to that found in tissues. Thus, this system also enables the researcher to distinguish between events that are caused by direct cell-cell contact and secreted factors.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Collagen Type V Antibody, Chemicon®, from rabbit
Sigma-Aldrich
Anti-Actin, α-Smooth Muscle - Cy3 antibody, Mouse monoclonal, clone 1A4, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-Collagen, Type I antibody produced in mouse, clone COL-1, ascites fluid