The aim of this project is to develop an efficient method for short term culture of circulating tumor cells (CTC) isolated from patients with liver and prostate cancer. Isolation will be performed using the reported assay ISET (isolation by size of epithelial tumor cells), since epithelial cells have a significantly larger size as compared to peripheral blood leukocytes. Filtration without using fixative buffer will allow to isolate living CTC.
The major challenge of this project is to develop an efficient method to stimulate growing of CTC. Tumor cells, in fact, stop growing when they circulate. The assay has been planned to avoid transfection of CTC with oncogenes and growth factors, since transfection would modify the CTC biological properties, including invasion potential and response to drugs. Several feeder layers, 3-dimensional media and organ culture will be used to help growing of tumor cells.
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