Methods of tissue generation and tissue engineered compositions

Abstract

Provided are methods and compositions for constructing stable mammalian embryonic epithelial tissues and organs as well as constructing kidney tissue, and treating renal failure. Disclosed are methods of using an active epithelial growth factor having the capability of effectuating induction of growth and morphogenesis is cells.

What is claimed is: 
     
         1 . A method of generating a tubular tissue structure, comprising:
 (a) contacting a stem cell with a cell survival agent or biological active agent to stimulate growth and proliferation;   (b) contacting the cells with a branching agent that promotes formation of tubular tissue branches and/or globular morphology to generate ureteric bud or Wolffian duct cell tissue;   (c) combining the ureteric bud tissue with metenephric mesenchyme in a biocompatible matrix; and   (d) culturing the combination to form a tubular tissue or kidney tissue in vitro.   
     
     
         2 . A method of generating a tubular tissue structure, comprising:
 (a) contacting a stem cell with a cell survival agent or biological active agent to stimulate growth and proliferation;   (b) contacting the cells with an agent that promotes formation of metenephric mesenchyme cells to generate a metenephric mesenchyme;   (c) combining the metenephric mesenchyme tissue with ureteric bud tissue in a biocompatible matrix; and   (d) culturing the combination to form a tubular tissue or kidney tissue.   
     
     
         3 . A method of generating a tubular tissue structure, comprising:
 (a) contacting a metenephric mesenchyme progenitor cell with a bioactive agent that induces growth and proliferation of the MM progenitor;   (b) contacting a ureteric bud or Wolffian duct progenitor cell with a bioactive agent that induces growth and proliferation of the UB or WD progenitor;   (c) growing the progenitor cells to form a population;   (d) inducing branching or the formation of globular structures in the UB or WD progenitor cells to generate a population of UBs or WDs by contacting the cells with a branching factor;   (e) inducing formation of MM from the MM progenitors by culturing the cells with a differentiation factor;   (f) combining the MM and UB and/or WD cells in a biocompatible matrix or gel; and   (d) culturing the combination to form a tubular tissue or kidney tissue.   
     
     
         4 . The method of  claim 1 ,  2 , or  3 , wherein the cell survival agent is selected from the group consisting of FGF1, FGF7 and a combination thereof either alone or in combination with one or more of GDNF, PTN, HRG, or BSN-CM, wherein the growth agent is selected from the group consisting of FGF1, FGF7, FGF1 and FGF7, PTN and GDNF, FGF1 and GDNF, FGF7 and GDNF, BSN-CM and FGF1, HRG and FGF1, PTN and FGF1, BSN and FGF7, HRG and FGF7, PTN and FGF7, BSN and FGF1 and GDNF, HRG and FGF1 and GDNF, PTN and FGF1 and GDNF, BSN and FGF7 and GDNF, HRG and FGF7 and GDNF, and PTN and FGF7 and GDNF. 
     
     
         5 . The method of  claim 1 ,  2 , or  3 , wherein the branching agent is selected from the group consisting of FGF1, FGF7, FGF1 and FGF7, PTN and GDNF, FGF1 and GDNF, FGF7 and GDNF, BSN-CM and FGF1, HRG and FGF1, PTN and FGF1, BSN and FGF7, HRG and FGF7, PTN and FGF7, BSN and FGF1 and GDNF, HRG and FGF1 and GDNF, PTN and FGF1 and GDNF, BSN and FGF7 and GDNF, HRG and FGF7 and GDNF, and PTN and FGF7 and GDNF. 
     
     
         6 . The method of  claim 1 ,  2 , or  3 , further comprising:
 culturing stem or progenitor cells in vitro under conditions that induce branching morphogenesis to generate a population of UBs, WDs, or UBs and WDs comprising tubular branches; and   subdividing the UB, WD, OR UB AND WD population; and   resuspending each subpopulation in culture media and repeating.   
     
     
         7 . A method for in vitro culturing and propagating ureteric bud, Wolffian duct bud, or ureteric and Wolffian duct bud tissue, comprising:
 isolating ureteric bud, Wolffian duct bud, or ureteric and Wolffian duct bud tissue from mesenchyme tissue obtained from embryonic kidney rudiments;   culturing the isolated ureteric bud, Wolffian duct bud, or ureteric and Wolffian duct bud tissue in a biocompatible matrix in the presence of a culture medium comprising a growth factor, growth factor combination as set forth in Table 2 for a sufficient time and under sufficient conditions to produce tubular branches within the biocompatible matrix;   separating the plurality of branch tips to generate bud fragments; and   culturing each of the bud fragments in a biocompatible matrix with a culture medium comprising pleiotrophin and/or heregulin or an active fragment thereof.   
     
     
         8 . The method of  claim 7 , wherein the biocompatible matrix comprises a material selected from the group consisting of a cotton, a collagen, a polyglycolic acid, a cat gut suture, a cellulose, a gelatin, a dextran, a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, a nitrocellulose compound, and a Matrigel. 
     
     
         9 . The method of  claim 8 , wherein the material is treated to contain proteoglycans, Type I collagen, Type IV collagen, laminin, proteoglycans, fibronectin, or combinations thereof. 
     
     
         10 . A method comprising:
 differentiating stem cells to form metenephric mesenchyme tissue;   differentiating stem cells to form ureteric bud cells; and   combining the UB cells and MM cells in a biocompatible matrix or gel; and   culturing the combination tissue to form a rudimentary kidney tissue.