Use of adipose tissue-derived stromal stem cells in treating fistula

Abstract

Provided herein are novel methods and compositions utilizing adipose tissue-derived stromal stem cells for treating fistulae.

1 - 35 . (canceled) 
     
     
         36 . A method of preparing a pharmaceutical composition, comprising thawing a cryopreserved composition and formulating the thawed composition with human serum albumin (HSA) and Dulbecco's Modified Eagle's Medium (DMEM) to produce the pharmaceutical composition, wherein the cryopreserved composition comprises an isolated cell population comprising adipose tissue-derived stromal stem cells, wherein the isolated cell population was ex vivo generated by culturing cells from adipose tissue for at least six days and passaging the cells from adipose tissue at least four times, wherein the concentration of the adipose tissue-derived stromal stem cells in the cryopreserved composition is at least about 5×10 6  cells/mL. 
     
     
         37 . The method of  claim 36 , wherein fewer than 15% of the isolated cell population in the cryopreserved composition express the CD34 marker as measured by flow cytometry and wherein at least 50% of the isolated cell population in the cryopreserved composition express the CD90 marker as measured by flow cytometry. 
     
     
         38 . The method of  claim 36 , wherein at least 85% of the adipose tissue-derived stromal stem cells in the cryopreserved composition express the CD44 marker and more than 50% of the human adipose-derived stromal stem cells express the CD29 and CD105 markers as measured by flow cytometry. 
     
     
         39 . The method of  claim 36 , wherein at least 95% of the adipose tissue-derived stromal stem cells in the cryopreserved composition express the CD44 marker as measured by flow cytometry. 
     
     
         40 . The method of  claim 36 , wherein at least 99% of the adipose tissue-derived stromal stem cells in the cryopreserved composition express the CD44 marker as measured by flow cytometry. 
     
     
         41 . The method of  claim 36 , wherein the adipose tissue-derived stromal stem cells were passaged at least six times. 
     
     
         42 . The method of  claim 36 , wherein the cryopreserved composition further comprises HSA. 
     
     
         43 . The method of  claim 36 , wherein the pharmaceutical composition further comprises an adhesive. 
     
     
         44 . The method of  claim 43 , wherein the adhesive comprises a fibrin glue or gel. 
     
     
         45 . The method of  claim 36 , wherein the pharmaceutical composition further comprises a therapeutic agent. 
     
     
         46 . The method of  claim 45 , wherein the therapeutic agent comprises an anti-inflammatory agent, an immunosuppressive agent, a biological agent, an antibiotic, an antidiarrheal agent or a combination thereof. 
     
     
         47 . A method of preparing a pharmaceutical composition, comprising formulating an isolated cell population with HSA and DMEM to produce the pharmaceutical composition, wherein the isolated cell population comprises adipose tissue-derived stromal stem cells, wherein the isolated cell population was ex vivo generated by culturing cells from adipose tissue for at least six days and passaging the cells from adipose tissue at least four times, wherein the concentration of the adipose tissue-derived stromal stem cells in the pharmaceutical composition is at least about 5×10 6  cells/mL. 
     
     
         48 . The method of  claim 47 , wherein fewer than 15% of the isolated cell population in the pharmaceutical composition express the CD34 marker as measured by flow cytometry and wherein at least 50% of the isolated cell population in the pharmaceutical composition express the CD90 marker as measured by flow cytometry. 
     
     
         49 . The method of  claim 47 , further comprising aliquoting the pharmaceutical composition into one or more doses of at least about 10×10 6  cells/dose. 
     
     
         50 . The method of  claim 47 , further comprising aliquoting the pharmaceutical composition into one or more doses of at least about 20×10 6  cells/dose.