High throughput bioreactor

Abstract

A continuous flow bioreactor system that includes a bioreactor, an optional post-bioreactor preparation chamber, a cell sorter, and an optional pre-bioreactor preparation chamber in a closed loop, useful for enriching a heterogeneous cell population growing in the bioreactor with an isolated subpopulation of cells.

What is claimed is:  
     
         1 . A bioreactor system comprising: 
 at least one bioreactor; and    at least one cell sorter coupled to the bioreactor;    wherein bioreactor and flow cytometer are in fluid communication with each other in a closed system.    
     
     
         2 . The bioreactor system of  claim 1 , further comprising at least one post-bioreactor preparation chamber coupled to the bioreactor and the cell sorter, wherein the bioreactor, the post-bioreactor preparation chamber and the cell sorter are in fluid communication with each other in the closed system.  
     
     
         3 . The bioreactor system of  claim 2  comprising a plurality of post-bioreactor preparation chambers in parallel or in series.  
     
     
         4 . The bioreactor system of  claim 2 , further comprising at least one pre-bioreactor preparation chamber coupled to the flow cytometer and the bioreactor, wherein the bioreactor, the post-bioreactor preparation chamber, the flow cytometer and the pre-bioreactor preparation chamber are in fluid communication with each other in the closed system.  
     
     
         5 . The bioreactor system of  claim 1 , further comprising at least one pre-bioreactor preparation chamber coupled to the flow cytometer and the bioreactor, wherein the bioreactor, the flow cytometer and the pre-bioreactor preparation chamber are in fluid communication with each other in the closed system.  
     
     
         6 . The bioreactor system of  claim 1  wherein the closed system maintains a sterile environment.  
     
     
         7 . The bioreactor system of  claim 1  wherein the cell sorter comprises a flow cytometer.  
     
     
         8 . The bioreactor system of  claim 7  wherein the flow cytometer comprises an optical flow cytometer.  
     
     
         9 . The bioreactor system of  claim 8  wherein the optical flow cytometer sorts cells based upon differences in light absorption, light scattering, or excitation.  
     
     
         10 . The bioreactor system of  claim 8  wherein the optical flow cytometer sorts cells based upon ultraviolet absorptions, fluorescence or phosphorescence.  
     
     
         11 . The bioreactor system of  claim 7  wherein the flow cytometer comprises an acoustic flow cytometer.  
     
     
         12 . The bioreactor system of  claim 1  wherein the cell sorter sorts cells based upon magnetic, electromagnetic, radiation, enzymatic or chemical properties of the cells.  
     
     
         13 . A bioreactor system comprising: 
 at least one bioreactor for providing an environment for growing cells; and    at least one cell sorter for separating the continuous stream of cells from the bioreactor into separate subpopulations, the cell sorter coupled to the bioreactor to return a separate subpopulation from the cell sorter to the bioreactor;    wherein the bioreactor and cell sorter are in fluid communication with each other in a closed system.    
     
     
         14 . A bioreactor system comprising: 
 at least one bioreactor for providing an environment for growing cells;    at least one post-bioreactor preparation chamber for preparing a continuous stream of cells from the bioreactor for separation, the post-bioreactor preparation chamber coupled to the bioreactor; and    at least one cell sorter for separating the continuous stream of cells from the bioreactor into separate subpopulations, the cell sorter coupled to the post-bioreactor separation chamber and the bioreactor to return a separate subpopulation from the cell sorter to the bioreactor;    wherein the bioreactor, post-bioreactor preparation chamber and cell sorter are in fluid communication with each other in a closed system.    
     
     
         15 . A bioreactor system comprising: 
 at least one bioreactor for providing an environment for growing cells; and    at least one cell sorter for separating the continuous stream of cells from the bioreactor into separate subpopulations, the cell sorter coupled to the bioreactor;    at least one pre-bioreactor preparation chamber for preparation a separated subpopulation of cells to be returned to the bioreactor, the pre-bioreactor preparation chamber coupled to the cell sorter and the bioreactor;    wherein the bioreactor, cell sorter and pre-bioreactor preparation chamber are in fluid communication with each other in a closed system.    
     
     
         16 . A bioreactor system comprising: 
 at least one bioreactor for providing an environment for growing cells;    at least one post-bioreactor preparation chamber for preparing a continuous stream of cells from the bioreactor for separation, the post-bioreactor preparation chamber coupled to the bioreactor;    at least one cell sorter for separating the continuous stream of cells prepared in the post-bioreactor separation chamber into separate subpopulations, the cell sorter coupled to the post-bioreactor separation chamber; and    at least one pre-bioreactor preparation chamber for preparation a separated subpopulation of cells to be returned to the bioreactor, the pre-bioreactor preparation chamber coupled to the cell sorter and the bioreactor;    wherein the bioreactor, post-bioreactor preparation chamber, cell sorter and pre-bioreactor preparation chamber are in fluid communication with each other in a closed system.    
     
     
         17 . The bioreactor system of any of claims  13 - 16  wherein the cell sorter comprises a flow cytometer.  
     
     
         18 . A method for accumulating cells of interest in a bioreactor, the method comprising: 
 growing a heterogeneous population of cells in one or more bioreactors, wherein the heterogenous population comprises at least one cell of interest;    removing a continuous stream of the heterogeneous population of cells from the bioreactor;    separating the heterogeneous population of cells in the continuous stream of cells into subpopulations of cells based on cell characteristics; and    returning a separated subpopulation of cells to the bioreactor to enrich the bioreactor for the separated subpopulation of cells, wherein the separated subpopulation of cells comprises at least one cell of interest.    
     
     
         19 . A method for accumulating cells of interest in a bioreactor, the method comprising: 
 growing a heterogenous population of cells in one or more bioreactors, wherein the heterogenous population comprises at least one cell of interest;    removing a continuous stream of the heterogeneous population of cells from the bioreactor;    preparing the continuous stream of cells for separation;    separating the heterogeneous population of cells in the continuous stream of cells into subpopulations of cells based on cell characteristics;    preparing a separated subpopulation of cells for return to the bioreactor; and    returning a separated subpopulation of cells to the bioreactor to enrich the bioreactor for the separated subpopulation of cells, wherein the separated subpopulation of cells comprises at least one cell of interest.    
     
     
         20 . The method of  claim 19  wherein preparing the continuous stream of cells for separation includes at least one activity selected from the group consisting of staining the cells, washing the cells and changing the concentration of the cells.  
     
     
         21 . The method of  claim 20  wherein preparing the continuous stream of cells for separation includes staining the cells.  
     
     
         22 . The method of  claim 19  wherein preparing the separated subpopulation of cells for return to the bioreactor includes at least one activity selected from the group consisting of washing the cells, filtering the cells and changing the concentration of the cells.  
     
     
         23 . The method of  claim 19  wherein the heterogenous population of cells is separated by analyzing at least one characteristic of the cells selected from the group consisting of light absorption, light scattering, excitation, ultraviolet absorption, fluorescence, phosphorescence, acoustic properties, magnetic properties, electromagnetic properties, radiation, enzymatic activity, affinity binding and other chemical properties of the cells.  
     
     
         24 . The method of  claim 19  wherein the heterogenous population of cells comprises cells selected from the group consisting of bacteria, protozoa, fungal cells, invertebrate cells, and vertebrate cells.  
     
     
         25 . The method of  claim 19  wherein the heterogenous population of cells comprises mammalian cells.  
     
     
         26 . The method of  claim 19  wherein the heterogenous population of cells comprises hybridoma cells, stem cells or hamster CHO cells.  
     
     
         27 . A method of using the bioreactor system of any of claims  1 ,  13 ,  14 ,  15  or  16  comprising: 
 growing a heterogenous population of cells in one or more bioreactors, wherein the heterogenous population comprises at least one cell of interest;  
 removing a continuous stream of the heterogeneous population of cells from the bioreactor;  
 preparing the continuous stream of cells for separation in a post-bioreactor separation chamber;  
 separating the heterogeneous population of cells in the continuous stream of cells into subpopulations of cells based on cell characteristics;  
 preparing a separated subpopulation of cells in a pre-bioreactor chamber for return to the bioreactor; and  
 returning a separated subpopulation of cells to the bioreactor to enrich the bioreactor for the separated subpopulation of cells, wherein the separated subpopulation of cells comprises at least one cell of interest.  
 
     
     
         28 . The method of  claim 27  wherein the cell sorter comprises a flow cytometer.