Methods of monitoring angiogenesis and metastasis in three dimensional co-cultures
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsChangge Fang, Enrique Zudaire, Frank Cuttitta
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Abstract
This disclosure relates to fluorescent cell lines and to the use of such cell lines in monitoring cellular activity, such as angiogenesis. This disclosure further relates to the use of such cell lines in a three-dimensional cell culture to monitor angiogenic and metastatic potential of tumor cells and selecting personalized therapeutics for treatment of cancer.
Claims21
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 tumor spheroid colony or a sample of a tumor biopsy, suspended in the solidified gel matrix;
- 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.
A method of selecting a personalized anti-angiogenic or anti-metastatic treatment for cancer in a subject comprising:
- preparing multiple three-dimensional co-cultures, each 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 tumor spheroid colony or a sample of a tumor biopsy, suspended in the solidified gel matrix;
- and a third layer on top of the second layer, comprising culture medium, wherein all but one of the co-cultures further comprises at least one test agent comprising an anti-angiogenic or anti-metastatic compound in the first, second, or third layers;
- incubating the three-dimensional co-cultures;
- detecting at least one of endothelial cell proliferation, endothelial cell tubule formation or tumor cell angiotropism of the cells in the second layer;
- and
- selecting the at least one test agent having the greatest effect on at least one of endothelial cell proliferation, endothelial cell tubule formation or tumor cell angiotropism in comparison to endothelial cell proliferation, endothelial cell tubule formation or tumor cell angiotropism in the cells of the co-culture without the test agent in the medium.
Dependencies are inferred from the claim text.