Cell composition and methods of making the same

Abstract

Provided herein are improved methods for the formulation of compositions comprising placental stem cells, and improved compositions and cell formulations produced thereby.

What is claimed is: 
     
         1 . A method of making a composition comprising cells, comprising:
 (a) contacting said cells with a solution comprising dextran and human serum albumin (HSA) to form a cell-containing solution;   (b) filtering the cell-containing solution;   (c) if said cell-containing solution comprises greater than about 10±3×10 6  cells per milliliter, diluting said cells to no more than about 10±3×10 6  cells per milliliter with a first dilution solution comprising dextran; and   (d) optionally diluting the cell-containing solution with a second dilution solution comprising dextran but not comprising HSA, thereby making a composition comprising cells.   
     
     
         2 . The method of  claim 1 , wherein said cells are cryopreserved after step (c) and before step (d). 
     
     
         3 . The method of  claim 1 , wherein said dextran in said first dilution solution or said second dilution solution is dextran 40. 
     
     
         4 . The method of  claim 1 , wherein said dextran in said first dilution solution and said second dilution solution is dextran 40. 
     
     
         5 . The method of  claim 1 , wherein said dextran 40 in said first dilution solution is 5.5% dextran 40. 
     
     
         6 . The method of  claim 1 , wherein said HSA in said solution comprising HSA is 10% HSA. 
     
     
         7 . The method of  claim 1 , wherein said first dilution solution comprises HSA. 
     
     
         8 . The method of  claim 7 , wherein said HSA in said first dilution solution is 10% HSA. 
     
     
         9 . The method of  claim 1 , wherein said first dilution solution further comprises a cryoprotectant. 
     
     
         10 . The method of  claim 9 , wherein said cryoprotectant is DMSO. 
     
     
         11 . The method of  claim 1 , wherein said dextran 40 in said second dilution solution is 10% dextran 40. 
     
     
         12 . The method of  claim 1 , wherein said solution in step (a) comprises a cryoprotectant. 
     
     
         13 . The method of  claim 1 , wherein said composition comprising cells comprises about 7.5% to about 9% dextran. 
     
     
         14 . The method of  claim 1 , wherein said composition comprising cells comprises about 1.0±0.3×10 6  cells per milliliter to about 5.0±1.5×10 6  cells per milliliter. 
     
     
         15 . The method of  claim 1 , wherein said cells are placental cells. 
     
     
         16 . The method of  claim 15 , wherein said stem cells are isolated human adherent placental cells. 
     
     
         17 . A method of making a composition comprising cells, comprising:
 (a) suspending a plurality of isolated human adherent placental cells in a 5.5% dextran 40, 10% HSA solution to form a cell-containing solution;   (b) filtering the cell-containing solution through a 70 μM filter;   (c) diluting the cell-containing solution in 5.5% dextran 40, 10% HSA, and 5% DMSO to no more than about 10±3×10 6  cells/mL;   (d) cryopreserving the cells;   (e) thawing the cells; and   (f) optionally diluting the cell-containing solution 1:1 to 1:11 with 10% dextran 40 to produce said composition comprising cells.   
     
     
         18 . A method of making a composition comprising cells, comprising:
 (a) centrifuging a plurality of isolated human adherent placental cells to collect the cells;   (b) resuspending the cells in 5.5% dextran 40;   (c) centrifuging the cells to collect the cells;   (d) resuspending the cells in a 5.5% dextran 40 solution that comprises 10% HSA to form a cell-containing solution;   (e) filtering the cell-containing solution through a 70 μM to 100 μM filter;   (f) diluting the cell-containing solution in 5.5% dextran 40, 10% HSA, and 5% DMSO to no more than about 10±3×10 6  cells/mL;   (g) cryopreserving the cells;   (h) thawing the cells; and   (i) optionally diluting the cell-containing solution 1:1 to 1:11 with 10% dextran 40 to produce said composition comprising cells.   
     
     
         19 . A method of making a composition comprising isolated human adherent placental cells, comprising:
 (a) providing a plurality of isolated human adherent placental cells in a solution comprising 5.5% dextran 40 and 10% human serum albumin (HSA) to form a solution comprising isolated human adherent placental cells;   (b) filtering said solution comprising isolated human adherent placental cells with filter that removes visible cell clumps to produce filtered isolated human adherent placental cells;   (c) diluting said filtered isolated human adherent placental cells with an amount of a solution comprising 5.5% dextran 40, 10% HSA and 5% dimethylsulfoxide (DMSO) sufficient to bring said filtered isolated human adherent placental cells to about 10±3×10 6  cells per milliliter; and   (d) diluting said isolated human adherent placental cells with 10% dextran 40 at a ratio of about 1:1 to about 1:11 isolated human adherent placental cells:dextran 40 to produce said composition.   
     
     
         20 . The method of  claim 1 , wherein said filter is a 70 μM filter. 
     
     
         21 . The method of  claim 1 , wherein said filter is a 100 μM filter. 
     
     
         22 . The method of  claim 1 , wherein said cells are isolated human adherent placental cells. 
     
     
         23 . The method of  claim 22  wherein said isolated human adherent placental cells are CD10 + , CD34 −  and CD105 + . 
     
     
         24 . The method of  claim 23 , wherein said CD10′ − , CD34 −  and CD105 +  cells are CD200 + . 
     
     
         25 . The method of  claim 24 , wherein said CD10 + , CD34 − , CD105 +  and CD200 +  cells are either CD45 −  or CD90 + . 
     
     
         26 . The method of  claim 24 , wherein said CD10 + , CD34 − , CD105 +  and CD200 +  cells are CD45 −  and CD90 + . 
     
     
         27 . A composition made by the method of  claim 1 . 
     
     
         28 . A method of making a composition comprising isolated human adherent placental cells, comprising contacting said isolated human adherent placental cells with a solution comprising 5.5% dextran and 10% HSA; filtering the isolated human adherent placental cells; and diluting said isolated human adherent placental cells to about 10±3×10 6  cells per milliliter with a solution comprising 10% dextran 40 (w/v). 
     
     
         29 . A composition comprising a plurality of isolated human adherent placental cells in a solution comprising 10% dextran 40, wherein said composition comprises between about 1.0±0.3×10 6  cells per milliliter to about 5.0±1.5×10 6  cells per milliliter, and wherein said composition comprises no macro cell clumps. 
     
     
         30 . The composition of  claim 29 , wherein said placental stem cells are thawed cryopreserved cells. 
     
     
         31 . The composition of  claim 29 , wherein said composition is contained within a bag. 
     
     
         32 . The composition of  claim 29 , wherein said composition comprises no macro cell clumps. 
     
     
         33 . The composition of  claim 29 , wherein said composition comprises fewer than about 200 micro cell clumps per 10 6  cells. 
     
     
         34 . The composition of  claim 29 , wherein said composition comprises fewer than about 150 micro cell clumps per 10 6  cells. 
     
     
         35 . The composition of  claim 29 , wherein said composition comprises fewer than about 100 micro cell clumps per 10 6  cells. 
     
     
         36 . The composition of  claim 29 , wherein said isolated human adherent placental cells are CD10 + , CD34 −  and CD105 + . 
     
     
         37 . The composition of  claim 36 , wherein said CD10 + , CD34 −  and CD105 +  cells are CD200 + . 
     
     
         38 . The composition of  claim 37 , wherein said CD10 + , CD34 − , CD105 +  and CD200 +  cells are either CD45 −  or CD90 + . 
     
     
         39 . The composition of  claim 37 , wherein said CD10 + , CD34 − , CD105 +  and CD200 +  cells are CD45 −  and CD90 + . 
     
     
         40 . The composition of  claim 29 , wherein said composition is a pharmaceutical composition. 
     
     
         41 . The method of  claim 1 , further comprising concentrating the composition comprising cells to about 5×10 6  cells per milliliter to 1×10 8  cells per milliliter. 
     
     
         42 . The method of  claim 41 , further comprising subcutaneously administering the composition comprising cells to a subject.