Polymeric Carriers and Methods

Abstract

Provided are methods of controlling disassociation of cells from a carrier, compositions, and methods of collecting cells. The methods of controlling disassociation of cells from a carrier may include contacting a polymeric carrier with one or more digesting agents to disassociate at least a portion of a plurality of cells from the polymeric carrier. The polymeric carrier may be crosslinked with a crosslinker including at least one of a redox sensitive moiety, a UV light sensitive moiety, a pH sensitive moiety, and a temperature sensitive moiety.

1 . A method of controlling disassociation of cells from a carrier, the method comprising:
 providing a polymeric carrier and a plurality of cells adhered to the polymeric carrier, wherein the polymeric carrier is a microcarrier; and   contacting the polymeric carrier with one or more digesting agents to disassociate at least a portion of the plurality of cells from the polymeric carrier;   wherein the polymeric carrier comprises gelatin, and is crosslinked with a crosslinker comprising at least one redox sensitive moiety, wherein the at least one redox sensitive moiety comprises a disulfide bond, and the crosslinker comprises a compound having the following structure:   
       
         
           
           
               
               
           
         
         wherein R 1  and R 2  are independently selected from a divalent C 1 -C 20  hydrocarbyl, and X and Y are independently selected from an epoxide or an N-hydroxysuccinimide ester moiety. 
       
     
     
         2 . The method of  claim 1 , wherein X and Y are the epoxide, and the crosslinker has the following structure: 
       
         
           
           
               
               
           
         
       
     
     
         3 . The method of  claim 2 , wherein R 1  and R 2  are unsubstituted divalent C 1 -C 5  hydrocarbyls, and the crosslinker has the following structure: 
       
         
           
           
               
               
           
         
       
     
     
         4 . The method of  claim 2 , wherein R 1  and R 2  are unsubstituted divalent C 2  hydrocarbyls, and the crosslinker comprises 1,2-bis(2-(oxiran-2-yl)ethyl)disulfane: 
       
         
           
           
               
               
           
         
       
     
     
         5 . The method of  claim 1 , wherein X and Y are the N-hydroxysuccinimide ester moiety, and the crosslinker has the following structure: 
       
         
           
           
               
               
           
         
       
     
     
         6 . The method of  claim 5 , wherein R 1  and R 2  are unsubstituted divalent C 1 -C 5  hydrocarbyls, and the crosslinker has the following structure: 
       
         
           
           
               
               
           
         
       
     
     
         7 . The method of  claim 5 , wherein R 1  and R 2  are unsubstituted divalent C 2  hydrocarbyls, and the crosslinker comprises bis(2,5-dioxopyrrolidin-1-yl)-3,3′-disulfanediyldipropanoate: 
       
         
           
           
               
               
           
         
       
     
     
         8 . The method of  claim 1 , wherein the plurality of cells comprises human Mesenchymal stem cells (hMSCs), endothelial cells, or a combination thereof. 
     
     
         9 . The method of  claim 1 , wherein the polymeric carrier further comprises dextran, hyaluronic acid, agarose, chitosan, alginate, polyvinyl alcohol, polyamino acid, calcium alginate, polyacrylate, polyethylene glycol, polyethylene imine, polyanhydride, or a combination thereof. 
     
     
         10 . The method of  claim 1 , wherein the polymeric carrier further comprises dextran, hyaluronic acid, agarose, chitosan, alginate, polyvinyl alcohol, polyamino acid, or a combination thereof. 
     
     
         11 . The method of  claim 1 , wherein the polymeric carrier is a polymeric carrier bead. 
     
     
         12 . The method of  claim 1 , wherein the one or more digesting agents comprises a reducing agent. 
     
     
         13 . The method of  claim 12 , wherein the reducing agent comprises dithiothreitol (DTT), cysteine, glutathione, or a combination thereof. 
     
     
         14 . The method of  claim 12 , wherein the reducing agent comprises a solution of dithiothreitol (DTT) having a concentration of about 5 mM to about 25 mM. 
     
     
         15 . The method of  claim 14 , wherein the solution of dithiothreitol further comprises sodium citrate, trypsin, or a combination thereof. 
     
     
         16 . The method of  claim 1 , wherein the one or more digesting agents comprises an enzyme. 
     
     
         17 . The method of  claim 16 , wherein the enzyme comprises collagenase, dispase-like protease, trypsin, urease, papain, bromelain, pancreatin, gelatinase (MMP2 and MMP9), or a combination thereof. 
     
     
         18 . The method of  claim 1 , wherein the one or more digesting agents comprises a reducing agent and an enzyme. 
     
     
         19 . The method of  claim 18 , wherein the enzyme comprises collagenase, dispase-like protease, trypsin, urease, papain, bromelain, pancreatin, gelatinase (MMP2 and MMP9), or a combination thereof. 
     
     
         20 . The method of  claim 1 , wherein the ratio of the crosslinker (mmol) to the polymeric carrier (g) is about 0.1 mmol:10 g to about 1.9 mmol:10 g.