Tissue Engineering of Tumor Stromal Microenvironment with Application to Cancer Cell Invasion

The overall goal of the following procedure is to generate a native matrix from fibroblasts for assessing tumor cell invasion. This is accomplished first by growing fibroblasts on plastic in a sorbic acid enriched media to generate the native matrix. In the second step, the native matrix is detached and released from the plastic growing surface, and then this matrix is seeded with tumor cells and lifted to the air liquid interface after a fixed period.

These 3D cultures are harvested for histological analysis of the tumor cell invasion. Ultimately, the effects of the native fibroblast matrix on tumor cell invasion can be evaluated by immuno histological analysis. So the advantage of this technique for generating 3D typic cultures is that we can assess native matrix produced by fibroblasts themselves in the absence of synthetic or foreign substrates, such as cross species type one collagen Begin by seeding 200, 000 fibroblasts per well in six well plates in fiberblast media, supplemented with freshly prepared IC acid.