Bioreactor system

Abstract

A three dimensional cell culture and bioreactor system is provided. The system comprises one or more cell culture chamber. Each cell culture chamber comprises an inlet port and an outlet port in fluid communication with the cell culture chamber. The cell culture chambers may be segregated or in fluid communication with one another. The systems may be used to conduct drug efficacy test, isolate certain cell types from a complex tissue sample of multiple cell types, allow for the ex vivo culturing of patient tissue samples to help guide the course of treatment, and conduct co-culture experiments.

What is claimed is: 
     
         1 . A method for testing the efficacy of pharmaceutical agents using three-dimensional cell culturing systems comprising;
 culturing a target cell population in a cell culture chamber of a three-dimensional (3D) cell culturing system;   exposing the target cell population in the cell culture chamber to a pharmaceutical agent for a defined period of time and under a defined set of exposure conditions; and   determining a cell viability or other measurable parameter for the target cell population, wherein the target cell population is responsive to the pharmaceutical agent if the cell viability or other measurable parameter of the target cell population is less than the cell viability or other measurable parameter of a control cell population;   wherein the cell culture chamber of the 3D cell culturing system comprises an inlet port and an outlet port that are in communication with an interior volume of the cell culture chamber, the interior volume of the cell culture chamber housing a cell-scaffold construct and the target cell population.   
     
     
         2 . The method of  claim 1 , wherein the target cell population is a cancer cell population, and the pharmaceutical agent is an anti-cancer agent. 
     
     
         3 . The method of  claim 2 , wherein the cancer cell population is obtained from a patient biopsy sample. 
     
     
         4 . The method of  claim 1 , wherein the 3D culture system further comprises two or more cell culture chambers. 
     
     
         5 . The method of  claim 4 , wherein the control cell population is cultured in one of the two or more cell culture chambers. 
     
     
         6 . The method of  claim 4 , wherein the two or more cell culture chambers are connected, and wherein the two or more cell culture chambers are separated by an impermeable membrane portion. 
     
     
         7 . The method of  claim 4 , wherein the two or more cell culture chambers are connected, and wherein the two or more cell culture chambers are separated by a permeable membrane portion that allows fluid communication between the cell chambers. 
     
     
         8 . The method of  claim 1 , wherein the cell-culture chamber further comprises an access port through which an analytical probe is inserted to access either the interior or exterior of the cell chamber. 
     
     
         9 . A method for isolating residual cell populations from tissue samples after exposure to pharmaceutical agents comprising;
 culturing a primary cell population derived from at least a portion of a subject biopsy sample in a cell culture chamber of a 3D cell culturing system, wherein the primary cell population comprises one or more cell types;   treating the primary cell population with a first dose of one or more pharmaceutical agents, wherein exposure to the one or more pharmaceutical agent results in an increased ratio of a first residual cell type relative to other cell types in the primary cell population; and   culturing the residual cell type;   wherein the cell culture chamber of the 3D cell culturing system comprises an inlet port and an outlet port that are in communication with an interior volume of the cell culture chamber, the interior volume of the cell culture chamber housing a cell-scaffold construct and the target cell population.   
     
     
         10 . The method of  claim 9 , further comprising treating the residual cell type with a second dose of the one or more pharmaceutical agents to obtain an enriched residual cell type. 
     
     
         11 . The method of  claim 9 , further comprising;
 treating the cultured residual cell type with a second dose of the one or more pharmaceutical agents, wherein the second dose of the one or more pharmaceutical agents is a higher does than the first dose;   determining an efficacy of the second dose based at least in part on a cell viability of the residual cell type after treatment with the second dose.   
     
     
         12 . The method of  claim 9 , further comprising;
 treating the cultured residual cell type with a second dose of one or more different pharmaceutical agents.   
     
     
         13 . The method of  claim 9 , wherein the residual cell type comprises tumor stem cells, and the one or more pharmaceutical agents are anti-cancer agents. 
     
     
         14 . The method of  claim 9 , wherein the cell-culture chamber further comprises an access port through which an analytical probe is inserted to access either the interior of the cell chamber. 
     
     
         15 . A method for determining appropriate dosage regimens for one or more pharmaceutical agents, comprising
 culturing, in a first cell culture chamber of a 3D cell culture system, a residual cell population obtained from a tissue sample of a subject in need of treatment using the method of claim;   culturing, in a second cell culture chamber of a 3D cell culture system, a primary cell population derived from a tissue sample from the subject;   sequentially treating the enriched residual cell population and the primary cell population with increasing doses of one or more pharmaceutical agents;   determining a cell viability of the enriched residual cell population and the primary cell population at each dose; and   selecting a final dose of the one or more pharmaceutical agents for administration to the subject, wherein the final dose of the one or more pharmaceutical agents is the dose that demonstrate the greatest decrease in cell viability of both the primary cell population and the enriched residual cell population;   wherein each cell culture chamber of the 3D cell culturing system comprises an inlet port and an outlet port in communication with an interior volume of the cell culture chamber, the interior volume of the culture chamber housing a cell-scaffold construct and the target cell population.   
     
     
         16 . The method of  claim 15 , wherein the tissue sample is a cancer tissue sample, and the one or more pharmaceutical agents are anti-cancer agents. 
     
     
         17 . The method of  claim 15 , wherein the first cell culture chamber and the second cell culture chamber are connected, and wherein the first cell culture chamber and the second cell culture chamber are separated by an impermeable membrane portion. 
     
     
         18 . The method of  claim 15 , wherein the two or more cell culture chambers are connected, and wherein the two or more cell culture chambers are separated by a permeable membrane portion that allows fluid communication between the cell culture chambers. 
     
     
         19 . The method of  claim 15 , wherein each cell-culture chamber further comprises an access port through which an analytical probe is inserted to access either the interior or exterior of the cell culture chamber. 
     
     
         20 . A method for monitoring and adjusting patient treatment regimens over a course of treatment comprising;
 culturing a patient cell population obtained from a diseased tissue of a patient in need of treatment in a cell culture chamber of a 3D cell culture system;   treating, prior to initiating or changing the treatment regimen, the patient cell population with one or more candidate pharmaceutical agents at one or more doses;   determining a cell viability or other measured parameter of the patient cell population for each candidate pharmaceutical agent at each dose; and   selecting a pharmaceutical agent and a dose of the pharmaceutical agent for the treatment regimen, wherein the pharmaceutical agent and the dose that demonstrates the greatest decrease in the cell viability or other measured parameter of the patient cell population;   wherein each cell culture chamber of the 3D cell culturing system comprises an inlet port and an outlet port in communication with an interior volume of the cell culture chamber, the interior volume of the cell culture chamber housing a cell-scaffold construct and the target cell population.   
     
     
         21 . The method of  claim 20 , the patient cell population comprises cancer cells. 
     
     
         22 . The method of  claim 20 , wherein the cell-culture chamber further comprises an access port through which an analytical probe is inserted to access either the interior or exterior of the cell culture chamber. 
     
     
         23 . A method for co-culturing cells comprising;
 culturing a first cell population in a first culture chamber of a 3D culture system, the first culture chamber comprising an inlet port and an outlet port through which a cell culture medium can flow, the inlet port connected to an opening in a first side of a cell culture chamber, and the outlet port connected to a second side on an opposing side of the cell culture chamber, and comprising an additional opening on a bottom side of the cell culture chamber;   culturing a second cell population in a second culture chamber of the 3D culture system, the second culture chamber comprising an inlet port and an outlet port through which a cell culture medium can flow, the inlet port connected to an opening in a first side of the cell culture chamber, and the outlet port connected to a second side on an opposing side of the cell culture chamber, and comprising an additional opening on a top side of the cell culture chamber;   wherein the bottom side of the first culture chamber and the top side of the second culture chamber are connected to one another, and wherein the first culture chamber and the second culture chamber are separated by a membrane portion.   
     
     
         24 . The method of  claim 23 , further comprising
 treating the first and second cell population with one or more pharmaceutical agents; and   determining an efficacy, a toxicity, or both of the one or more pharmaceutical agents on the first and second cell populations by determining a cell viability for the first and second cell populations.   
     
     
         25 . The method of  claim 23 , wherein the first cell population is a diseased cell population isolated from a biological organism, and the second cell population is a disease free cell population isolated from the biological organism. 
     
     
         26 . The method of  claim 25 , wherein the diseased cell population is a cancer cell population. 
     
     
         27 . The method of  claim 23 , wherein the first cell culture chamber and the second cell culture chamber are connected, and wherein the first cell culture chamber and the second cell culture chamber are separated by an impermeable membrane portion. 
     
     
         28 . The method of  claim 23 , wherein the first cell culture chamber and the second cell culture chamber are connected and stacked together, and wherein the first and second cell culture chambers are separated by a permeable membrane portion that allows fluid communication between the two or more cell culture chambers. 
     
     
         29 . The method of  claim 23 , wherein each cell-culture chamber further comprises an access port through which an analytical probe is inserted to access either the interior or exterior of the cell culture chamber.