Bioreactors with multiple chambers

Abstract

A bioreactor for cultivating living cells in a liquid medium. In one embodiment of the present invention, the bioreactor has a first substrate having a first surface and an opposite second surface, defining a chamber therebetween for receiving the cells and the liquid medium. The bioreactor further has a barrier dividing the chamber into a first subchamber and a second subchamber, wherein the barrier has a porosity to allow the first subchamber and the second subchamber in fluid communication and allow at least one predetermined type of cells to permeate between the first subchamber and the second subchamber.

1 - 32 . (canceled) 
     
     
         33 . A bioreactor for cultivating living cells in a liquid medium comprising:
 (a) a substrate having a first surface and an opposite second surface, defining a chamber therebetween for receiving the cells and the liquid medium, wherein the chamber is formed with a center and a boundary;   (b) a first barrier enclosing the center and a portion of the chamber to form a central chamber;   (c) a second barrier positioned between the first barrier and the boundary so as to form an intermediate chamber and an outer chamber; and   (d) means for electrochemical measurements of the cells responsive to the liquid medium in at least one of the outer chamber, the intermediate chamber and the central chamber;   (e) means for optical measurements of the cells responsive to the liquid medium in at least one of the outer chamber, the intermediate chamber, and the central chamber,
 wherein the means for electrochemical measurements is positioned between and in communication with the substrate and the means for optical measurements, and wherein the first barrier has a first porosity to allow the central chamber and the intermediate chamber in fluid communication and allow at least a first predetermined type of cells to permeate between the central chamber and the intermediate chamber, and the second barrier has a second porosity to allow the outer chamber and the intermediate chamber in fluid communication and allow at least a second predetermined type of cells to permeate between the outer chamber and the intermediate chamber. 
   
     
     
         34 . The bioreactor of  claim 33 , wherein the central chamber is adapted for receiving a first type of material, the intermediate chamber is adapted for receiving a second type of material, and the outer chamber is adapted for receiving a third type of material. 
     
     
         35 . The bioreactor of  claim 34 , wherein each of the first type of material, the second type of material and the third type of material contains at least one selected from the group of cells, chemicals, and fluids. 
     
     
         36 . The bioreactor of  claim 33 , wherein the first predetermined type of cells comprises tumor cells, which normally is received in the central chamber corresponding to a tumor space. 
     
     
         37 . The bioreactor of  claim 36 , wherein the second predetermined type of cells comprises normal tissue cells, which normally is received in the intermediate chamber corresponding to a tissue space. 
     
     
         38 . The bioreactor of  claim 37 , wherein the outer chamber is corresponding to a vascular space adapted for receiving endothelial cells, macrophage cells, neutophil cells, any combination of them, or other immune cell type. 
     
     
         39 . The bioreactor of  claim 33 , further comprising a biocompatible coating layer applied to the chamber walls at the boundary. 
     
     
         40 . The bioreactor of  claim 39 , wherein the biocompatible coating layer comprises a material that may inhibit cell adhesion to the biocompatible coating layer, enhance cell adhesion to the biocompatible coating layer, or function as a fluorescent marker or indicator of the state of cells. 
     
     
         41 . The bioreactor of  claim 33 , further comprising at least one inlet or outlet port and an input or output transfer channel in fluid communication with the inlet or outlet port and the external chamber for allowing delivery of the cells, fluids or chemicals to the outer chamber. 
     
     
         42 . The bioreactor of  claim 41 , further comprising at least one inlet or outlet port and an input or output transfer channel in fluid communication with the inlet or outlet port and the central chamber for allowing delivery of the cells, fluids or chemicals to the central chamber. 
     
     
         43 . The bioreactor of  claim 42 , further comprising at least one inlet or outlet port and an input or output transfer channel in fluid communication with the inlet or outlet port and the intermediate chamber for allowing delivery of the cells, fluids or chemicals to the intermediate chamber. 
     
     
         44 . The bioreactor of  claim 33 , wherein the substrate is fabricated from glass, Mylar, PDMS, silicon, a polymer, a semiconductor, or any combination of them. 
     
     
         45 . The bioreactor of  claim 33 , wherein the first barrier comprises a porous material. 
     
     
         46 . The bioreactor of  claim 45 , wherein the first barrier is microfabricated so as to form a first structure allowing the fluid communication between the central chamber and the intermediate chamber. 
     
     
         47 . The bioreactor of  claim 46 , wherein the second barrier comprises a porous material. 
     
     
         48 . The bioreactor of  claim 47 , wherein the second barrier is microfabricated so as to form a second structure allowing the fluid communication between the outer chamber and the intermediate chamber, and the second structure is different from the first structure. 
     
     
         49 . (canceled) 
     
     
         50 . The bioreactor of  claim 33 , wherein the means for electrochemical measurements comprises:
 (i) a reference electrode;   (ii) a counter electrode; and   (iii) a plurality of individually addressable working electrodes.   
     
     
         51 . The bioreactor of  claim 50 , wherein the liquid medium comprises at least one analyte, and wherein the plurality of individually addressable working electrodes comprise a first group of individually addressable working electrodes adapted for sensing the concentration of a single analyte of the liquid medium at multiple locations in the outer chamber or the concentrations of a plurality of analytes of the liquid medium at multiple locations in the outer chamber at a time period shorter than a characterization reaction time related to at least one of cellular physiological activities of the cells. 
     
     
         52 . The bioreactor of  claim 51 , wherein the first group of individually addressable working electrodes are further adapted for measuring the metabolic variables related to the cells responsive to the liquid medium at multiple locations in the outer chamber at a time period shorter than a characterization reaction time related to at least one of cellular physiological activities of the cells. 
     
     
         53 . The bioreactor of  claim 52 , wherein the plurality of individually addressable working electrodes comprise a second group of individually addressable working electrodes adapted for sensing the concentration of a single analyte of the liquid medium at multiple locations in the central chamber or the concentrations of a plurality of analytes of the liquid medium at multiple locations in the central chamber at a time period shorter than a characterization reaction time related to at least one of cellular physiological activities of the cells. 
     
     
         54 . The bioreactor of  claim 53 , wherein the second group of individually addressable working electrodes are further adapted for measuring the metabolic variables related to the cells responsive to the liquid medium at multiple locations in the central chamber at a time period shorter than a characterization reaction time related to at least one of cellular physiological activities of the cells. 
     
     
         55 . The bioreactor of  claim 54 , wherein the plurality of individually addressable working electrodes comprise a third group of individually addressable working electrodes adapted for sensing the concentration of a single analyte of the liquid medium at multiple locations in the intermediate chamber or the concentrations of a plurality of analytes of the liquid medium at multiple locations in the intermediate chamber at a time period shorter than a characterization reaction time related to at least one of cellular physiological activities of the cells. 
     
     
         56 . The bioreactor of  claim 55 , wherein the third group of individually addressable working electrodes are further adapted for measuring the metabolic variables related to the cells responsive to the liquid medium at multiple locations in the intermediate chamber at a time period shorter than a characterization reaction time related to at least one of cellular physiological activities of the cells. 
     
     
         57 . The bioreactor of  claim 33 , wherein the first barrier is substantially circular. 
     
     
         58 . The bioreactor of  claim 33 , wherein the second barrier is substantially circular. 
     
     
         59 . The bioreactor of  claim 33 , wherein the boundary is substantially circular. 
     
     
         60 . A bioreactor for cultivating living cells in a liquid medium comprising:
 (a) a substrate having a first surface and an opposite second surface, defining a chamber therebetween for receiving the cells and the liquid medium with a boundary;   (b) means for dividing the chamber into plurality of subchambers;   (c) means for electrochemical measurements of the cells responsive to the liquid medium in at least one of the plurality of subchambers; and   (d) means for optical measurements of the cells responsive to the liquid medium in at least one of the plurality of subchambers,
 wherein the means for electrochemical measurements is positioned between and in communication with the substrate and the means for optical measurements, and wherein each of the plurality of subchambers is in fluid communication with at least another one of the plurality of subchambers. 
   
     
     
         61 . The bioreactor of  claim 60 , wherein the dividing means comprises a barrier to divide the chamber into a first subchamber and a second subchamber, and wherein the barrier has a porosity to allow the first subchamber and the second subchamber in fluid communication and allow at least one predetermined type of cells to permeate between the first subchamber and the second subchamber. 
     
     
         62 . The bioreactor of  claim 60 , wherein the dividing means comprises a first barrier and a second barrier to divide the chamber into a first subchamber, a second subchamber and a third subchamber, and wherein the first barrier has a first porosity to allow the first subchamber and the intermediate subchamber in fluid communication and at least a first predetermined type of cells to permeate between the first subchamber and the second subchamber, and the second barrier has a second porosity to allow the second subchamber and the third subchamber in fluid communication and at least a second predetermined type of cells to permeate between the second subchamber and the third subchamber. 
     
     
         63 . The bioreactor of  claim 62 , wherein the first porosity and the second porosity can be same or different. 
     
     
         64 . The bioreactor of  claim 60 , wherein the dividing means comprises a plurality of n barriers, n being an integer greater than zero, to divide the chamber into n+1 subchambers. 
     
     
         65 . The bioreactor of  claim 64 , wherein each of n barriers has a corresponding porosity that can be same or different from that of other barriers.