In vitro engineered, regenerated urinary tract tissue compositions and methods for producing same

Abstract

A method for providing a urinary tract tissue graft composition includes providing a tissue culture frame and a segment of small intestinal submucosa and positioning the segment of small intestinal submucosa in the tissue culture frame such that the segment of small intestinal submucosa is suspended and held in a taut position by the tissue culture frame. Smooth muscle and urothelial cells are isolated from a tissue specimen of a subject and cultured, and then seeded upon the segment of small intestinal submucosa, thereby forming a urinary tract tissue graft. A tissue culture frame in which such a urinary tract tissue graft may be formed is also disclosed.

1 . A urinary tract tissue graft composition, comprising a segment of small intestinal submucosa having at least one cell type seeded on a surface of the segment of small intestinal submucosa, the at least one cell type selected from the group consisting of smooth muscle cells and urothelial cells.  
     
     
         2 . The urinary tract tissue graft composition of  claim 1  wherein the at least one cell type exhibits three dimensional growth and matrix penetrance.  
     
     
         3 . A urinary tract tissue graft composition, comprising a segment of small intestinal submucosa having smooth muscle cells and urothelial cells seeded on a surface of the segment of small intestinal submucosa.  
     
     
         4 . The urinary tract tissue graft composition of  claim 3  wherein the smooth muscle cells and urothelial cells exhibit three dimensional growth and matrix penetrance.  
     
     
         5 . The urinary tract tissue graft composition of  claim 3 , wherein the smooth muscle cells are seeded on a mucosal surface of the segment of small intestinal submucosa.  
     
     
         6 . The urinary tract tissue graft composition of  claim 5 , wherein the urothelial cells are seeded on a serosal surface of the segment of small intestinal submucosa.  
     
     
         7 . The urinary tract tissue graft composition of  claim 5 , wherein the urothelial cells are seeded upon the smooth muscle cells attached to the mucosal surface of the segment of small intestinal submucosa.  
     
     
         8 . The urinary tract tissue graft composition of  claim 3 , wherein the smooth muscle cells are bladder smooth muscle cells and the urothelial cells are bladder urothelial cells.  
     
     
         9 . A urinary tract tissue graft composition, comprising: 
 a segment of small intestinal submucosa having a mucosal surface and a serosal surface;    smooth muscle cells seeded on the mucosal surface of the segment of small intestinal submucosa; and    urothelial cells seeded on the serosal surface of the segment of small intestinal submucosa.    
     
     
         10 . The urinary tract tissue graft composition of  claim 9  wherein the smooth muscle cells and urothelial cells exhibit three dimensional growth and matrix penetrance.  
     
     
         11 . The urinary tract tissue graft composition of  claim 9 , wherein the smooth muscle cells are bladder smooth muscle cells and the urothelial cells are bladder urothelial cells.  
     
     
         12 . A urinary tract tissue graft composition, comprising: 
 a segment of small intestinal submucosa having a mucosal surface and a serosal surface;    smooth muscle cells seeded on the mucosal surface of the segment of small intestinal submucosa; and    urothelial cells seeded on the smooth muscle cells attached to the mucosal surface of the segment of small intestinal submucosa.    
     
     
         13 . The urinary tract tissue graft composition of  claim 12  wherein the smooth muscle cells and urothelial cells exhibit three dimensional growth and matrix penetrance.  
     
     
         14 . The urinary tract tissue graft composition of  claim 12 , wherein the smooth muscle cells are bladder smooth muscle cells and the urothelial cells are bladder urothelial cells.  
     
     
         15 . A method for repairing a damaged urinary tract tissue of a subject, comprising the steps of: 
 isolating and culturing smooth muscle cells from a urinary tract tissue specimen of a subject;    isolating and culturing urothelial cells from the urinary tract tissue specimen of the subject;    providing a tissue culture frame;    providing a segment of small intestinal submucosa having a mucosal surface and a serosal surface;    positioning the segment of small intestinal submucosa in the tissue culture frame such that the segment of small intestinal submucosa is suspended and held in a taut position by the tissue culture frame;    seeding the smooth muscle cells on the segment of small intestinal submucosa; and    seeding the urothelial cells on the segment of small intestinal submucosa;    allowing the segment of small intestinal submucosa having the smooth muscle cells and urothelial cells thereon to mature in culture such that the smooth muscle cells and urothelial cells exhibit three dimensional growth and matrix penetrance;    removing the segment of small intestinal submucosa from the tissue culture frame; and    contacting the damaged urinary tract tissue with the small intestinal submucosa under conditions such that growth of the urinary tract tissue occurs and the damaged urinary tract tissue is repaired, thereby restoring urological function.    
     
     
         16 . The method of  claim 15  wherein, in the steps of isolating and culturing smooth muscle cells from a urinary tract tissue specimen of a subject and isolating and culturing urothelial cells from the urinary tract tissue specimen of the subject, the urinary tract tissue specimen is bladder tissue, and the smooth muscle cells are bladder smooth muscle cells and the urothelial cells are bladder urothelial cells.  
     
     
         17 . The method of  claim 15  wherein, in the step of seeding the smooth muscle cells on the segment of small intestinal submucosa, the smooth muscle cells are seeded on the mucosal surface of the segment of small intestinal submucosa.  
     
     
         18 . The method of  claim 17  wherein, in the step of seeding the urothelial cells on the segment of small intestinal submucosa, the urothelial cells are seeded on the serosal surface of the segment of small intestinal submucosa.  
     
     
         19 . The method of  claim 17  wherein, in the step of seeding the urothelial cells on the segment of small intestinal submucosa, the urothelial cells are seeded on the smooth muscle cells attached to the mucosal surface of the segment of small intestinal submucosa.  
     
     
         20 . A method for repairing a damaged urinary tract tissue of a subject, comprising the steps of: 
 isolating and culturing smooth muscle cells from a urinary tract tissue specimen of a subject;    isolating and culturing urothelial cells from the urinary tract tissue specimen of the subject;    providing a tissue culture frame;    providing a segment of small intestinal submucosa having a mucosal surface and a serosal surface;    positioning the segment of small intestinal submucosa in the tissue culture frame such that the segment of small intestinal submucosa is suspended and held in a taut position by the tissue culture frame;    seeding smooth muscle cells on the mucosal surface of the segment of small intestinal submucosa; and    seeding urothelial cells on the serosal surface of the segment of small intestinal submucosa;    allowing the segment of small intestinal submucosa having the smooth muscle cells and urothelial cells thereon to mature in culture such that the smooth muscle cells and urothelial cells exhibit three dimensional growth and matrix penetrance;    removing the segment of small intestinal submucosa from the tissue culture frame; and    contacting the damaged urinary tract tissue with the small intestinal submucosa under conditions such that growth of the urinary tract tissue occurs and the damaged urinary tract tissue is repaired, thereby restoring urological function.    
     
     
         21 . The method of  claim 20  wherein, in the steps of isolating and culturing smooth muscle cells from a urinary tract tissue specimen of a subject and isolating and culturing urothelial cells from the urinary tract tissue specimen of the subject, the urinary tract tissue specimen is bladder tissue, and the smooth muscle cells are bladder smooth muscle cells and the urothelial cells are bladder urothelial cells.  
     
     
         22 . A method for repairing a damaged urinary tract tissue of a subject, comprising the steps of: 
 isolating and culturing smooth muscle cells from a urinary tract tissue specimen of a subject;    isolating and culturing urothelial cells from the urinary tract tissue specimen of the subject;    providing a tissue culture frame;    providing a segment of small intestinal submucosa having a mucosal surface and a serosal surface;    positioning the segment of small intestinal submucosa in the tissue culture frame such that the segment of small intestinal submucosa is suspended and held in a taut position by the tissue culture frame;    seeding smooth muscle cells on the mucosal surface of the segment of small intestinal submucosa; and    seeding urothelial cells on the smooth muscle cells attached to the serosal surface of the segment of small intestinal submucosa;    allowing the segment of small intestinal submucosa having the smooth muscle cells and urothelial cells thereon to mature in culture such that the smooth muscle cells and urothelial cells exhibit three dimensional growth and matrix penetrance;    removing the segment of small intestinal submucosa from the tissue culture frame; and    contacting the damaged urinary tract tissue with the small intestinal submucosa under conditions such that growth of the urinary tract tissue occurs and the damaged urinary tract tissue is repaired, thereby restoring urological function.    
     
     
         23 . The method of  claim 22  wherein, in the steps of isolating and culturing smooth muscle cells from a urinary tract tissue specimen of a subject and isolating and culturing urothelial cells from the urinary tract tissue specimen of the subject, the urinary tract tissue specimen is bladder tissue, and the smooth muscle cells are bladder smooth muscle cells and the urothelial cells are bladder urothelial cells.  
     
     
         24 . A tissue culture frame in which a membrane may be suspended and held in a taut position, thereby maintaining tension on the membrane such that cells may be seeded on the membrane.  
     
     
         25 . The tissue culture frame of  claim 24  wherein the membrane is a segment of small intestinal submucosa.  
     
     
         26 . A kit, comprising: 
 means for suspending a segment of small intestinal submucosa and holding the segment of small intestinal submucosa in a taut position such that cells may be seeded thereon; and    a segment of small intestinal submucosa suspended and held in a taut position such that cells may be seeded thereon.    
     
     
         27 . A kit, comprising: 
 a tissue culture frame in which a membrane may be suspended and held in a taut position, thereby maintaining tension on the membrane such that cells may be seeded on the membrane; and    a segment of small intestinal submucosa suspended in the tissue culture frame and held taut such that cells may be seeded thereon.