Fibrin microbeads and uses thereof

Abstract

The present invention provides fibrin microbeads that are biologically active and comprise extensively cross-linked fibrin(ogen), and a method for preparing the fibrin microbeads. The present invention also provides a composition comprising cells bound to the fibrin microbeads, and methods for culturing and separating cells using the fibrin microbeads of the present invention. Finally, the present invention provides methods for transplanting cells and engineering tissue using the fibrin microbeads of the present invention.

1 - 49 . (Canceled)  
     
     
         50 . A wound healing composition comprising wound healing promoting cells and fibrin microbeads, wherein the fibrin microbeads are prepared in the absence of glutaraldehyde as a cross-linking agent, and wherein the fibrin microbeads are prepared by a method comprising the sequential steps of: (i) preparing an aqueous solution comprising fibrinogen, thrombin and Factor XIII; (ii) prior to the onset of coagulation, contacting said aqueous solution with an oil heated to a temperature of about 50-80° C. to form an emulsion; (iii) mixing the emulsion at a temperature of about 50-80° C. until fibrin microbeads comprising extensively cross-linked fibrin(ogen) are obtained; and (iv) isolating the fibrin microbeads.  
     
     
         51 . The composition of  claim 50 , wherein the wound healing promoting cells are selected from the group consisting of fibroblasts, endothelial cells, chondrocytes, bone forming cells, cartilage forming cells, and combinations thereof.  
     
     
         52 . The composition of  claim 50 , wherein the fibrin microbeads comprise a bioactive agent.  
     
     
         53 . The composition of  claim 50 , wherein the fibrin microbeads comprise at least one bioactive agent selected from the group consisting of wound healing promoting agents, growth factors, glucocorticosteroids, steroids, antibiotics, antibacterial compounds, antiviral compounds, and antifungal compounds.  
     
     
         54 . The composition of  claim 50 , wherein the wound is a surgical wound, a burn, an ulcer, and/or a laceration.  
     
     
         55 . The composition of  claim 50 , wherein the composition is affixed to the wound using fibrin glue.  
     
     
         56 . The composition of  claim 50 , wherein at least 30% of the fibrin(ogen) is cross-linked.  
     
     
         57 . The composition of  claim 50 , wherein at least 50% of the fibrin(ogen) is cross-linked.  
     
     
         58 . The composition of  claim 50 , wherein the fibrin microbeads have a diameter of about 50-200 microns.  
     
     
         59 . The composition of  claim 50 , wherein the fibrin microbeads are biodegradable.  
     
     
         60 . A tissue forming composition comprising tissue forming cells and fibrin microbeads, wherein the fibrin microbeads are prepared in the absence of glutaraldehyde as a cross-linking agent, and wherein the fibrin microbeads are prepared by a method comprising the sequential steps of: (i) preparing an aqueous solution comprising fibrinogen, thrombin and Factor XIII; (ii) prior to the onset of coagulation, contacting said aqueous solution with an oil heated to a temperature of about 50-80° C. to form an emulsion; (iii) mixing the emulsion at a temperature of about 50-80° C. until fibrin microbeads comprising extensively cross-linked fibrin(ogen) are obtained; and (iv) isolating the fibrin microbeads.  
     
     
         61 . The composition of  claim 60 , wherein the tissue forming cells are selected from the group consisting of fibroblasts, smooth muscle cells, endothelial cells, chondrocytes, bone forming cells, cartilage forming cells, neuroblastoma cells, kidney cells, liver cells, pancreatic cells, thyroid cells, glial cells, and combination thereof.  
     
     
         62 . The composition of  claim 60 , wherein the fibrin microbeads comprise a bioactive agent.  
     
     
         63 . The composition of  claim 60 , wherein the fibrin microbeads comprise at least one bioactive agent selected from the group consisting of wound healing promoting agents, growth factors, glucocorticosteroids, steroids, antibiotics, antibacterial compounds, antiviral compounds, and antifungal compounds.  
     
     
         64 . The composition of  claim 60 , wherein the composition is affixed to a prosthetic device.  
     
     
         65 . The composition of  claim 60 , wherein the composition is affixed to a prosthetic device using fibrin glue.  
     
     
         66 . The composition of  claim 60 , wherein at least 30% of the fibrin(ogen) is cross-linked.  
     
     
         67 . The composition of  claim 60 , wherein at least 50% of the fibrin(ogen) is cross-linked.  
     
     
         68 . The composition of  claim 60 , wherein the fibrin microbeads have a diameter of about 50-200 microns.  
     
     
         69 . The composition of  claim 60 , wherein the fibrin microbeads are biodegradable.