Patent ID: 8679836

Claim:
A method for monitoring angiogenic or metastatic potential of tumor cells comprising: preparing a three-dimensional co-culture comprising: a first layer comprising a neutral polysaccharide polymer gel in contact with the bottom of a culture dish; a second layer on top of the first layer, comprising: a solidified gel matrix; endothelial cells dispersed in the solidified gel matrix; and tumor cells comprising either a single monoclonal tumor spheroid colony or a sample of a tumor biopsy, suspended in the solidified gel matrix, wherein the monoclonal tumor spheroid colony is produced using a method comprising: (i) preparing a culture in which monoclonal tumor spheroid colonies are grown, comprising: a bottom layer comprising 1% agarose with 20% FBS, 2× Antibiotic-Antimycotic, and 1×RPMI1640; and a top layer overlaying the bottom layer, wherein the top layer comprises isolated tumor cells suspended in 0.2% of agarose, 2× Antibiotic-Antimycotic, 20% FBS, and 1×RPMI1640; (ii) incubating the culture to grow monoclonal spheroid colonies; (iii) harvesting monoclonal tumor spheroid colonies from the culture; and (iv) resuspending the monoclonal tumor spheroids in phosphate buffered saline with 1% glucose, 0.3 mM EDTA, 0.5% BSA and 1× Antibiotic-Antimycotic; and a third layer on top of the second layer, comprising culture medium; incubating the three-dimensional co-culture; and detecting at least one of endothelial cell proliferation, endothelial cell tubule formation, or tumor cell angiotropism of the cells in the second layer that correlates in a tumor-specific manner with endothelial cell proliferation, endothelial cell vessel formation, or tumor cell angiotropism in vivo.