Engineered Three-Dimensional Breast Tissue, Adipose Tissue, and Tumor Disease Model

Abstract

Described are three-dimensional, engineered, biological breast tissues, adipose tissues, and tumor models, including breast cancer models.

1 - 30 . (canceled) 
     
     
         31 . A three-dimensional, engineered, biological breast cancer model comprising:
 a. breast stromal tissue, the stromal tissue comprising human mammary fibroblasts, human endothelial cells, and human adipocytes; and   b. breast cancer tumor tissue; the tumor tissue comprising breast cancer cells and human endothelial cells, the tumor tissue surrounded on all sides by the stromal tissue to form the three-dimensional, engineered, biological breast cancer model;   provided that the stromal tissue was bioprinted from a stromal bio-ink, the tumor tissue was bioprinted from a tumor bio-ink, or both the stromal tissue and the tumor tissue were bioprinted from their respective bio-inks.   
     
     
         32 . An array of three-dimensional, engineered, biological breast cancer models, each breast cancer model comprising:
 a. stromal tissue, the stromal tissue comprising human mammary fibroblasts, human endothelial cells, and human adipocytes; and   b. tumor tissue; the tumor tissue comprising breast cancer cells and human endothelial cells, the tumor tissue surrounded on all sides by the stromal tissue to form each three-dimensional, engineered, biological breast cancer model;   provided that the stromal tissue, the tumor tissue, or both the stromal tissue and the tumor tissue were bioprinted; provided that the array is adapted for use in a high throughput assay.   
     
     
         33 . A method of fabricating a three-dimensional, engineered, biological breast cancer model, the method comprising:
 a. preparing a stromal bio-ink, the stromal bio-ink comprising a plurality of stromal cell types, the stromal cell types comprising: an extrusion compound, human mammary fibroblasts, human endothelial cells, and human adipocytes;   b. preparing a tumor bio-ink, the tumor bio-ink comprising: an extrusion compound and a breast cancer cell type;   c. depositing the stromal bio-ink and the tumor bio-ink such that the tumor bio-ink is embedded in the stromal bio-ink and in contact with the stromal bio-ink on all sides; and   d. maturing the deposited bio-ink in a cell culture media to remove the extrusion compound and allow the cells to cohere to form a three-dimensional, engineered, biological breast cancer model.   
     
     
         34 . The method of  claim 33 , wherein depositing the stromal bio-ink and the tumor bio-ink further comprises:
 a. depositing a first sheet of stromal bio-ink on a surface;   b. depositing a continuous border of stromal bio-ink on the first sheet of stromal bio-ink to define a compartment, the compartment open on one side;   c. depositing a node of tumor bio-ink in the compartment; and   d. depositing a second sheet of stromal bio-ink to close the open side of the compartment.   
     
     
         35 . A method of identifying a therapeutic agent for cancer in an individual, the method comprising:
 a. preparing a stromal bio-ink, the stromal bio-ink comprising a plurality of stromal cell types;   b. preparing a tumor bio-ink, the tumor bio-ink comprising primary cancer cells from the individual;   c. depositing the stromal bio-ink and the tumor bio-ink such that the tumor bio-ink is embedded in the stromal bio-ink and in contact with the stromal bio-ink on all sides;   d. maturing the deposited bio-ink in a cell culture media to allow the cells to cohere to form a three-dimensional, engineered, biological construct;   e. applying a candidate therapeutic agent to the construct;   f. measuring viability of the cancer cells; and   g. selecting a therapeutic agent for the individual based on the measured viability of the cancer cells;   provided that at least one component of the construct was deposited by bioprinting.   
     
     
         36 . The method of  claim 35 , wherein depositing the stromal bio-ink and the tumor bio-ink further comprises:
 a. depositing a first sheet of stromal bio-ink on a surface;   b. depositing a continuous border of stromal bio-ink on the first sheet of stromal bio-ink to define a compartment, the compartment open on one side;   c. depositing a node of tumor bio-ink in the compartment; and   d. depositing a second sheet of stromal bio-ink to close the open side of the compartment.   
     
     
         37 . A method of identifying a therapeutic agent for breast cancer in an individual, the method comprising:
 a. preparing a breast stromal bio-ink, the stromal bio-ink comprising a plurality of breast stromal cell types;   b. preparing a breast tumor bio-ink, the tumor bio-ink comprising primary breast cancer cells from the individual;   c. depositing the stromal bio-ink and the tumor bio-ink such that the tumor bio-ink is embedded in the stromal bio-ink and in contact with the stromal bio-ink on all sides;   d. maturing the deposited bio-ink in a cell culture media to allow the cells to cohere to form a three-dimensional, engineered, breast cancer tumor model;   e. applying a candidate therapeutic agent to the breast cancer tumor model;   f. measuring viability of the breast cancer cells; and   g. selecting a therapeutic agent for the individual based on the measured viability of the breast cancer cells;   provided that the stromal bio-ink and the tumor bio-ink were deposited by bioprinting.   
     
     
         38 . The method of  claim 37 , wherein depositing the stromal bio-ink and the tumor bio-ink further comprises:
 a. depositing a first sheet of stromal bio-ink on a surface;   b. depositing a continuous border of stromal bio-ink on the first sheet of stromal bio-ink to define a compartment, the compartment open on one side;   c. depositing a node of tumor bio-ink in the compartment; and   d. depositing a second sheet of stromal bio-ink to close the open side of the compartment.