Viable cell compositions, and methods related to same

Abstract

Disclosed are viable cell compositions and related methods of preparation, maintenance and use. The viable cell composition can contain specified levels of cells, hydroxyethyl starch, and dimethylsulfoxide, and can be cryopreserved. The cryopreserved form of the composition can be thawed and combined with an aqueous liquid diluting medium to prepare a diluted viable cell composition that can contain specified, reduced levels of the dimethylsulfoxide and hydroxyethyl starch. The diluting medium can contain trehalose. The prepared, diluted viable cell composition can be administered to a patient.

1 . (canceled) 
     
     
         2 . A method for preparing viable cells for treating a patient, comprising:
 thawing a cryopreserved composition that includes viable cells and an aqueous cryopreservation medium, wherein the cryopreservation medium of the cryopreserved composition contains dimethylsulfoxide at a concentration of about 1% to 3% and hydroxyethyl starch at a concentration of about 2% to 10%, to provide a thawed viable cell composition having the viable cells suspended in an aqueous liquid; and   combining an aqueous liquid diluting medium with the thawed viable cell composition, to provide a diluted viable cell composition containing hydroxyethyl starch and dimethylsulfoxide, wherein the dimethylsulfoxide is present in the diluted viable cell composition at a concentration of about 0.5% or less.   
     
     
         3 . The method of  claim 2 , also comprising, prior to the thawing:
 storing a cryopreservation container in a freezer at a temperature in the range of about −60° C. to −100° C., the cryopreservation container containing the cryopreserved composition.   
     
     
         4 . The method of  claim 2 , wherein the thawed viable cell composition has a concentration of the viable cells in the range of about 1 million cells/ml to about 20 million cells/ml, and wherein the diluted viable cell composition has a concentration of the viable cells in the range of about 200,000 cells/ml to about 5 million cells/ml. 
     
     
         5 . The method of  claim 2 , also comprising administering the diluted viable cell composition to a patient by delivering the diluted viable cell composition into a blood vessel of the patient. 
     
     
         6 . The method of  claim 2 , wherein the cryopreservation medium of the cryopreserved composition contains the dimethylsulfoxide at a concentration of about 1.5% to 2.5%. 
     
     
         7 . The method of  claim 2 , wherein said volume ratio is at least 8:1, and wherein the dimethylsulfoxide is present in the diluted viable cell composition at a concentration of about 0.3% or less. 
     
     
         8 . The method of  claim 2 , wherein the cryopreservation medium of the cryopreserved composition contains the dimethylsulfoxide at a concentration of about 1.75% to 2.25%. 
     
     
         9 . The method of  claim 2 , wherein the viable cells comprise viable mesenchymal stem cells. 
     
     
         10 - 14 . (canceled) 
     
     
         15 . The method of  claim 2 , wherein the diluted viable cell composition contains the hydroxyethyl starch at a concentration of at least about 0.1%. 
     
     
         16 . The method of  claim 2 , wherein the diluted viable cell composition contains the DMSO at a concentration in the range of about 0.05% to about 0.5% and the hydroxyethyl starch at a concentration in the range of about 0.1% to about 6%. 
     
     
         17 . The method of  claim 2 , wherein the aqueous liquid diluting medium comprises trehalose. 
     
     
         18 . The method of  claim 17 , wherein the trehalose is present in the aqueous liquid diluting medium at a concentration of about 1% to about 10%. 
     
     
         19 . The method of  claim 18 , wherein the trehalose is present in the diluted viable cell composition at a concentration of about 2% to about 7%. 
     
     
         20 . The method of  claim 2 , wherein the cryopreserved composition contains the dimethylsulfoxide at a concentration of about 2% and the hydroxyethyl starch at a concentration of about 6%. 
     
     
         21 . A method for maintaining viable cells, comprising:
 maintaining in a cryopreserved condition a composition that includes viable cells and an aqueous cryopreservation medium, wherein the cryopreservation medium contains dimethylsulfoxide at a concentration of about 1% to 3% and hydroxyethyl starch at a concentration of about 2% to 10%.   
     
     
         22 . The method of  claim 28 , wherein said maintaining comprises maintaining the composition at a temperature in the range of about −60° C. to −100° C. 
     
     
         23 - 26 . (canceled) 
     
     
         27 . The method of  claim 21 , wherein the viable cells comprise viable mesenchymal stem cells. 
     
     
         28 . (canceled) 
     
     
         29 . The method of  claim 21 , wherein the composition contains the dimethylsulfoxide at a concentration of about 2% and the hydroxyethyl starch at a concentration of about 6%. 
     
     
         30 . A method for treating a patient with viable cells, comprising:
 administering a viable cell composition to the patient, the viable cell composition containing viable cells suspended in an aqueous liquid containing dimethylsulfoxide at a concentration of 0.5% or less and hydroxyethyl starch at a concentration that is greater than that of the dimethylsulfoxide and that is at least 0.1%.   
     
     
         31 . The method of  claim 30 , wherein said administering comprises delivering the viable cell composition into a blood vessel of the patient. 
     
     
         32 . The method of  claim 30 , wherein the viable cells comprise viable mesenchymal stem cells. 
     
     
         33 . (canceled) 
     
     
         34 . The method of  claim 30 , wherein the viable cell composition also comprises trehalose. 
     
     
         35 . The method of  claim 34 , wherein the trehalose is present in the viable cell composition at a concentration of about 2% to about 7%. 
     
     
         36 . The method of  claim 30 , also comprising, prior to said administering, preparing the viable cell composition by combining an aqueous liquid diluting medium with a starting viable cell composition containing dimethylsulfoxide at a concentration of about 1 to 3% and hydroxyethyl starch at a concentration of about 2 to 10%. 
     
     
         37 . The method of  claim 36 , wherein said combining is conducted with the aqueous liquid diluting medium and starting viable cell composition in a volume ratio of at least 5:1. 
     
     
         38 . The method of  claim 36 , wherein the aqueous liquid diluting medium comprises trehalose. 
     
     
         39 . A product, comprising:
 a cryopreservation container containing a cryopreserved composition that includes viable cells and an aqueous cryopreservation medium, wherein the cryopreservation medium of the cryopreserved composition contains dimethylsulfoxide at a concentration of about 1% to 3% and hydroxyethyl starch at a concentration of about 2% to 10%.   
     
     
         40 - 42 . (canceled) 
     
     
         43 . A viable cell composition for administration to a patient, comprising:
 an aqueous medium comprising dimethylsulfoxide at a concentration of less than 0.5%, hydroxyethyl starch at a concentration of at least about 0.1%; and   viable cells suspended in the aqueous medium at a concentration of about 200,000 cells/ml to about 20 million cells/ml.   
     
     
         44 . The viable cell composition of  claim 43 , wherein:
 the aqueous medium also comprises trehalose.   
     
     
         45 - 47 . (canceled)