Selective expansion of target cell populations

Abstract

The invention provides a method of selective expansion of a predetermined target population of cells, including (a) introducing a starting sample of cells into a growth medium; (b) causing cells of the predetermined target cell population to divide; and (c) contacting the starting cells with a selection element, comprising a plurality of selective binding molecules with specific affinity either for target cells or for non-target cells to select cells of said predetermined target population from other cells in the growth medium. The invention also provides systems for selective expansion and methods of treating patients with cell populations and products.

1 . A method of selective expansion of a predetermined target population of cells, said method comprising: 
 introducing a starting sample of cells into a growth medium;    causing cells of said predetermined target cell population to divide; and    contacting the cells in the growth medium with a selection element, comprising a plurality of selective binding molecules with specific affinity either for target cells or for a first population of non-target cells, so as to select cells of said predetermined target population from other cells in the growth medium.    
     
     
         2 . The method of  claim 1  wherein said selective binding molecules are specific for said target cells.  
     
     
         3 . The method of  claim 1  wherein said selective binding molecules are specific for non-target cells.  
     
     
         4 . The method of  claim 2 , further comprising contacting the starting cells with a reverse selection element comprising selective binding molecules with specific affinity for a second population of non-target cells.  
     
     
         5 . The method of  claim 1 , further comprising removing cells from said growth medium.  
     
     
         6 . The method of  claim 5 , wherein the removed cells are cells of said predetermined target cell population.  
     
     
         7 . The method of  claim 5  wherein the removed cells are non-target cells.  
     
     
         8 . The method of  claim 1  wherein said starting sample of cells includes said target cells, and said expansion is clonogenic.  
     
     
         9 . The method of  claim 1  wherein said starting sample of cells includes progenitors of said target cells.  
     
     
         10 . The method of  claim 1 , further comprising removing the growth medium prior to said contacting step.  
     
     
         11 . The method of  claim 1 , wherein said selection element comprises a solid support to which said selective binding molecules are bound.  
     
     
         12 . The method of  claim 11  wherein said growth medium is disposed in or caused to flow through a chamber.  
     
     
         13 . The method of  claim 12  further comprising causing said growth medium to recycle through said chamber, flowing from an inlet, through said chamber, to an outlet of said chamber, and returning from the outlet to the inlet via a conduit.  
     
     
         14 . The method of  claim 2  wherein said selective binding molecules bind to a cell surface antigen on cells of said predetermined target cell population but not on cells not in said target cell population.  
     
     
         15 . The method of  claim 14  wherein said selective binding molecule is a biotinylated antibody specific for an antigen on the surfaces of said cells of said predetermined target cell population.  
     
     
         16 . The method of  claim 3  wherein said selective binding molecules bind to a cell surface antigen that is on cells of the non-target cell population but not on cells of said target cell population.  
     
     
         17 . The method of  claim 11  further comprising the step of causing plasma to bind to regions of said solid support on which selective binding molecules are not present.  
     
     
         18 . The method of  claim 17  wherein said plasma is autologous plasma.  
     
     
         19 . The method of  claim 17  wherein said plasma is a type-matched allogeneic plasma.  
     
     
         20 . The method of  claim 11 , further comprising the step of contacting the solid support with an agent that is capable of binding to regions of said solid support on which said selective binding molecules are not present to prevent non-specific interaction between said regions and materials contacting said solid support.  
     
     
         21 . The method of  claim 1  wherein said growth medium is conditioned by cells present in the growth medium.  
     
     
         22 . The method of  claim 28  further comprising causing the growth medium to recycle through a fluid flow loop, thereby enhancing the level of conditioning.  
     
     
         23 . The method of  claim 29  further comprising concurrently removing dividing cells from the fluid flow loop during recycling.  
     
     
         24 . The method of  claim 1 , further comprising regulating the oxygen saturation of the growth medium to be from 0% to 20% relative to the solubility of oxygen in said fluid at equilibrium with air at 37° C. and 1 atm pressure.  
     
     
         25 . A system for continuous selective expansion of a predetermined target population of cells, comprising: 
 a growth medium for supporting cell division;    a chamber for receiving said growth medium; and    a selection element, comprising a plurality of binding sites bearing a selective binding molecule having a specific affinity for cells of said predetermined target cell population, positioned to contact said growth medium during or after cell division.    
     
     
         26 . A system for continuous selective expansion of a predetermined target population of cells, comprising: 
 a growth medium for causing cell division;    a chamber for receiving said growth medium; and    a selection element, comprising a plurality of binding sites bearing a selective binding molecule having a specific affinity for cells of cell populations other than said predetermined target cell population and having substantially less affinity for cells of said predetermined target cell population, said selection element being positioned to contact said growth medium during or after cell division.    
     
     
         27 . A method of selective expansion of a predetermined target population of cells, said method comprising: 
 introducing fluid containing a plurality of cells into a growth medium;    causing cells of said predetermined target cell population to divide; and    selecting cells of said predetermined target population from other cells in the growth medium;    wherein said causing and selecting steps are carried out substantially simultaneously.    
     
     
         28 . A method of enhancing the immune function of a patient, said method comprising introducing a sample containing relatively undifferentiated cells into a growth medium; 
 causing said relatively undifferentiated cells to divide;    selecting relatively undifferentiated cells from other cell populations present in said growth medium; and    administering a portion of the selected relatively undifferentiated cells to the patient.    
     
     
         29 . The method of  claim 28  wherein the sample is autologous.  
     
     
         30 . The method of  claim 29  wherein the sample comprises blood or bone marrow of said patient.  
     
     
         31 . The method of  claim 28  wherein said sample is allogeneic.  
     
     
         32 . A method of treating a patient in need of cell infusion, comprising: 
 introducing a sample containing relatively undifferentiated cells into a growth medium;    introducing into said growth medium one or more growth factors selected to cause said relatively undifferentiated cells to differentiate to form a predetermined population of cells;    causing cells in said growth medium to divide;    selecting cells of said predetermined population from other cells present in said growth medium; and    administering cells of said predetermined population to the patient.    
     
     
         33 . The method of  claim 32  wherein said second population of cells comprises lymphoid precursors, myeloid precursors, or erythroid precursors.  
     
     
         34 . A method of reducing negative feedback during selective clonogenic expansion of relatively undifferentiated cells comprising immobilizing a population of relatively undifferentiated cells on a solid support, providing a nutrient-containing medium to said cells, continuously removing waste products of said cells from contact with said immobilized, relatively undifferentiated cells, while continuously removing relatively undifferentiated cells immediately after they are formed.  
     
     
         35 . A method of treating a patient in need of a therapeutic compound, said method comprising: 
 (a) providing a sample containing a first population of cells which produce said therapeutic compound;    (b) passing a portion of the sample through a chamber containing a solid support capable of supporting a plurality of living cells adhered thereto, wherein said support comprises a plurality of binding sites with specific affinity for said first population of cells so that a portion of said cells bind to said binding sites;    (c) causing medium containing nutrients to flow through said chamber to cause said bound cells to produce said compound;    (d) harvesting said compound from the chamber; and    (e) administering said compound to the patient.    
     
     
         36 . A method of making a bioreactor for continuous selective clonogenic expansion of a predetermined population of cells, said method comprising: 
 (a) providing a chamber having an inlet, an outlet, and a passage for fluid flow from the inlet to the outlet, and, disposed in the chamber, a solid support having a plurality of activated sites;    (b) binding a selective binding molecule with specific affinity for said predetermined population of cells to the activated sites to enable the activated sites to support a plurality of living cells adhered thereto;    (c) introducing into the chamber an agent which is capable of binding to regions of said solid support on which said selective binding molecules are not present and preventing non-specific interaction between said regions and materials subsequently introduced into the chamber;    (d) causing fluid containing a plurality of cells of said cell population to flow through said chamber, so that a portion of said cells bind to said binding sites; and    (e) introducing a growth medium to the chamber.    
     
     
         37 . The method of  claim 36  wherein said agent is plasma.  
     
     
         38 . The method of  claim 37  wherein said plasma is autologous with respect to the cell population.  
     
     
         39 . The method of  claim 37  wherein said plasma is a type-matched allogenic plasma.