Process for producing a tissue transplant construct for reconstructing a human or animal organ

Abstract

A process for producing a tissue transplant construct for reconstructing a human or animal organ. The process may include the steps of: (a) isolation and two-dimensional cultivation of organ-specific tissue cells; (b) application of the organ-specific tissue cells to a biocompatible, collagen-containing membrane; and, (c) cultivation of the organ-specific tissue cells on the membrane with biochemical and mechanical stimulation of the organ-specific tissue cells. Tissue transplant constructs and methods for using tissue transplant constructs are also taught.

1 . A process for fabricating a urinary bladder tissue, comprising the steps of:
 providing an acellular tissue membrane obtained from one or more of urinary bladder, urinary bladder submucosa, aorta, urethra, small intestinal submucosa or dermal matrix, the acellular tissue membrane having a first side and an opposing second side;   collecting smooth muscle cells and interstitial fibroblastic cells from a urinary bladder;   collecting urothelial cells from a urinary bladder;   culturing smooth muscle cells and interstitial fibroblastic cells in two dimensions to confluence in a plurality of passages;   culturing the urothelial cells in two dimensions to confluence in a plurality of passages;   seeding the first side of the acellular tissue membrane with smooth muscle cells and interstitial fibroblastic cells from a selected passage;   seeding the second side of the acellular tissue membrane with urothelial cells from a selected passage; and,   applying tension cyclically to the seeded acellular tissue membrane, wherein the periodicity of the tension, and the strain resulting therefrom, is regular, and wherein the tension is cyclically applied under cell culturing conditions including a serum-conditioned culture medium, and resulting in a urinary bladder tissue having at least one layer of smooth muscle cells and interstitial fibroblastic cells, and at least one layer of urothelial cells bounded by the acellular tissue membrane.   
     
     
         2 . The process of  claim 1 , wherein the acellular tissue membrane is predominantly composed of collagen. 
     
     
         3 . The process of  claim 1 , wherein the steps of collecting cells from a urinary bladder further comprises collecting such cells from the same species into which the fabricated tissue is to be implanted. 
     
     
         4 . The process of  claim 1 , wherein the steps of seeding the acellular tissue membrane further comprises selecting a tissue culture passage from passage two to passage four. 
     
     
         5 . The process of  claim 1 , wherein the step of applying tension cyclically further comprises applying sufficient tension to result in up to 10% strain of the acellular tissue membrane. 
     
     
         6 . The process of  claim 1 , wherein the step of applying tension cyclically further comprises a stretching period of about 10 seconds to about 20 seconds, and a relaxing period of about 10 seconds. 
     
     
         7 . The process of  claim 6 , wherein the step of cyclically applying tension continues until the acellular tissue membrane comprises confluently cultured cells. 
     
     
         8 . The process of  claim 1 , wherein the serum-conditioned culture medium comprises autologous serum, or fetal bovine serum conditioned by proliferating urothelial cells. 
     
     
         9 . The process of  claim 1 , wherein the serum-conditioned culture medium comprises about 5% serum for a first cultivation period, 1% serum for a second cultivation period, and about 0% serum for a third cultivation period. 
     
     
         10 . The process of  claim 9 , wherein the first cultivation period is about 14 days, the second cultivation period is about 13 days, and the third cultivation period is about three days. 
     
     
         11 . A fabricated urinary bladder tissue made by the process of  claim 10 .