Muscle Derived Cells for the Treatment of Urinary Tract Pathologies and Methods of Making and Using the Same

Abstract

The present invention provides muscle-derived progenitor cells that show long-term survival following transplantation into body tissues and which can augment soft tissue following introduction (e.g. via injection, transplantation, or implantation) into a site of soft tissue. Also provided are methods of isolating muscle-derived progenitor cells, and methods of genetically modifying the cells for gene transfer therapy. The invention further provides methods of using compositions comprising muscle-derived progenitor cells for the augmentation and bulking of mammalian, including human, soft tissues in the treatment of various functional conditions, including malformation, injury, weakness, disease, or dysfunction. In particular, the present invention provides treatments and amelioration for urinary incontinence and other urinary tract pathologies.

We claim: 
     
         1 . A method for preparing a cell population containing muscle derived cells (MDCs) useful for administration to treat urinary tract disease in a mammalian subject, comprising:
 (a) suspending cells isolated from human skeletal muscle in a first cell culture container for a duration sufficient to adhere a first cell population to the container and to leave a second cell population remaining unadhered and in a culture medium in the container;   (b) transferring the culture medium and second cell population from the first cell culture container to a second cell culture container;   (c) allowing cells from the second cell population to attach to the second cell culture container; and   (d) isolating the cells attached to the second cell culture container to obtain said cell population containing MDCs.   
     
     
         2 . A method for preparing a cell population containing muscle derived progenitor cells (MDCs), comprising:
 mincing skeletal muscle tissue obtained from a human patient to be treated with muscle derived progenitor cells, wherein the skeletal muscle tissue has been stored under storage conditions causing stress to the skeletal muscle tissue;   digesting the minced muscle tissue to obtain a mixed population of cells; and   preparing an enriched population of muscle derived progenitor cells from the mixed population of cells.   
     
     
         3 . The method of  claim 2 , wherein said storage conditions include cooling. 
     
     
         4 . The method of  claim 2 , wherein said storage conditions include storing the skeletal muscle tissue in hypothermic medium. 
     
     
         5 . The method of  claim 4 , wherein the hypothermic medium contains an antibiotic. 
     
     
         6 . The method of  claim 4 , wherein said storage conditions include storing the skeletal muscle tissue at a temperature from about 4° C. to room temperature. 
     
     
         7 . The method of  claim 2 , wherein the enriched population of muscle derived progenitor cells is prepared based on differential adherence characteristics of cells of the mixture of cells. 
     
     
         8 . The method of  claim 7 , wherein said preparing an enriched population of muscle derived progenitor cells includes suspending said mixture of cells in a first cell culture container for a period sufficient for a first population of cells to adhere to the container and a second population of cells to remain non-adhered to the container, and transferring the second population of cells to a second cell culture container. 
     
     
         9 . The method of  claim 2 , also comprising freezing the enriched population of muscle derived progenitor cells in the presence of cryopreservation medium. 
     
     
         10 . The method of  claim 2 , wherein the skeletal muscle tissue has been stored for 1 to 7 days under storage conditions causing stress to the skeletal muscle tissue 
     
     
         11 . A method of treating a urinary tract disease in a human patient in need thereof, comprising administering to the urinary tract of the patient an enriched population of muscle derived progenitor cells prepared in accordance with  claim 2 .