Methods and Systems for T Cell Expansion

Abstract

The present disclosure provides a system for mimicking the secondary lymphoid organs where suspension cells (e.g., T cells) are expanded; methods of expanding, activating, and transfecting the suspension cells in the synthetic microenvironment, and suspension cells produced by such systems and methods.

1 . A system for expanding, activating, and/or transfecting suspension cells comprising:
 a three-dimensional functionalized porous microcarrier; and   suspension cells.   
     
     
         2 . (canceled) 
     
     
         3 . The system of  claim 1 , wherein the suspension cells are T cells. 
     
     
         4 .- 7 . (canceled) 
     
     
         8 . The system of  claim 1 , wherein the functionalized porous microcarrier comprises at least one of antibodies, aptamers, and phage-display identified peptide ligands. 
     
     
         9 . The system of  claim 1 , wherein the functionalized porous microcarrier comprises antibodies that are specific for the suspension cells. 
     
     
         10 . The system of  claim 3 , wherein the functionalized porous microcarrier comprises antibodies that are specific for T cells. 
     
     
         11 . The system of  claim 1 , wherein the functionalized porous microcarrier comprises one or more of anti-CD2, anti-CD3 and anti-CD28 antibodies. 
     
     
         12 .- 13 . (canceled) 
     
     
         14 . The system of  claim 1  further comprising an open bioreactor comprising a static culture vessel with a gas-permeable bottom. 
     
     
         15 . The system of  claim 1  further comprising a closed bioreactor selected from the group consisting of a stirred-type bioreactor, a bag bioreactor, a perfusion bioreactor, and combinations thereof. 
     
     
         16 . A system for expanding, activating, and/or transfecting T cells comprising:
 a three-dimensional functionalized macroporous microcarrier;   T cells; and   at least one of:
 a culture medium; 
 a cytokine; 
 a viral vector; and 
 a growth factor. 
   
     
     
         17 . The system of  claim 16  comprising at least the cytokine selected from the group consisting of IL2, IL7, IL15, and combinations thereof. 
     
     
         18 . The system of  claim 16 , wherein the T cells are selected from the group consisting of recombinant T cells, gene modified T cells, chimeric antigen receptor (CAR) T cells, unmodified T cells, CCR7+CD62+ central memory T cells, and combinations thereof. 
     
     
         19 . The system of  claim 16  comprising at least the viral vector that is configured for use in gene therapy. 
     
     
         20 . The system of  claim 16  comprising at least the viral vector comprising a CAR transgene, the system further comprising an additional gene selected from the group consisting of a therapeutic gene, surface marker gene, reporter gene, suicide genes, chemokine receptor gene, cytokine-expressing gene, immune-checkpoint receptor gene, and combinations thereof. 
     
     
         21 . (canceled) 
     
     
         22 . A method comprising:
 obtaining a blood sample from a patient;   isolating suspension cells from the blood sample;   introducing at least a portion of the suspension cells to a bioreactor comprising a three-dimensional functionalized porous microcarrier;   activating at least a portion of the suspension cells introduced into the bioreactor; and   expanding at least a portion of the activated suspension cells.   
     
     
         23 .- 28 . (canceled) 
     
     
         29 . The method of  claim 22 , wherein the porous microcarrier comprises one or more of proteins, carbohydrates, lipids and nucleic acids. 
     
     
         30 .- 31 . (canceled) 
     
     
         32 . The method of  claim 22 , wherein the functionalized porous microcarrier comprises at least one of antibodies, aptamers, and phage-display identified peptide ligands. 
     
     
         33 .- 34 . (canceled) 
     
     
         35 . The method of  claim 22 , wherein the functionalized porous microcarrier comprises one or more of anti-CD2, anti-CD3 and anti-CD28 antibodies. 
     
     
         36 .- 47 . (canceled) 
     
     
         48 . (canceled) 
     
     
         49 . A suspension cell obtained by the method of  claim 22 . 
     
     
         50 .- 56 . (canceled) 
     
     
         57 . A pharmaceutical composition comprising:
 at least one suspension cell of  claim 49 ;   at least one carrier; and   at least one additional therapeutic agent;   wherein the composition is formulated for intravenous administration.   
     
     
         58 .- 63 . (canceled) 
     
     
         64 . The method of of  claim 22  further comprising:
 transfecting at least a portion of the expanded suspension cells; 
 preparing at least a portion of the transfected suspension cells for transfusion into the patient; and 
 transfusing at least a portion of the prepared suspension cells into the patient; 
 wherein the porous microcarrier is configured for activation, expansion, and/or transfection of the suspension cells. 
 
     
     
         65 .- 68 . (canceled)