Macrotumor engineering
AssigneePurdue Research Foundation
InventorsSophie Andree Lelievre, Rahim Rahimi, Tim Kwok
Life of this patent
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Abstract
A collagen-fibronectin-based method enables the production of tumors of centimeter size with recognizable pathological traits. The method supports reproducing tumor heterogeneity with preinvasive and invasive phenotypes and stacking of tumor portions using a paper-scaffold with 80 μm-punched holes for larger nodule creation and easy separation of tumor portions for analysis. Macrotumors are convenient for testing drug delivery and therapeutic tools that necessitate a minimum tumor size relevant to in vivo.
Claims10
A multilayer stacking for culturing macrotumors, wherein the multilayer stacking comprises a plurality of cell culture amenable substrates, wherein each substrate has a plurality of pores with a diameter of 60-100 μm, wherein the density of the pores on each substrate is 2-8 pores/mm2, and wherein each substrate is pre-coated with a solution comprising one or more extracellular matrix.
A method of preparing a microtumor, comprising
- a) providing a pellet comprising a plurality of microtumors of 50-800 microns;
- b) providing a first cell culture amenable substrate, wherein the first substrate has a plurality of pores with a diameter of 60-100 μm, wherein the density of the pores on each substrate is 2-8 pores/mm2, and the first substrate is pre-coated with a solution comprising one or more extracellular matrix, and depositing the pellet comprising a plurality of microtumors onto the first cell culture amenable substrate;
- c) adding an extracellular matrix solution onto the pellet deposited on the first substrate to allow the pellet to be embedded by the extracellular matrix solution;
- d) providing a second cell culture amenable substrate, wherein the second substrate has a plurality of pores with a diameter of 60-100 μm, wherein the density of the pores on each substrate is 2-8 pores/mm2, and the second substrate is pre-coated with a solution comprising one or more extracellular matrix, and placing the second cell culture amenable substrate onto the extracellular matrix embedded microtumors deposited on the first cell culture amenable substrate;
- e) adding a solution comprising one or more extracellular matrix solution onto the pellet of deposited on the second substrate to allow the pellet to be embedded by the extracellular matrix solution;
- f) optionally repeating steps a)-e) to provide a total of 2-5 cell culture amenable substrates, wherein one pellet comprising a plurality of microtumors is deposited between two adjacent substrates;
- g) incubating the obtained multilayer stacking in a cell culture medium.
Dependencies are inferred from the claim text.