Use of perfusion decellularized liver for islet cell recellularization

Abstract

The invention provides a method to prepare a graft comprising a recellularized extracellular matrix of a mammalian liver, liver lobe or portion thereof, and a method of using the recellularized extracellular matrix of a mammalian liver, liver lobe or portion thereof.

1 . A method to prepare a graft comprising a recellularized extracellular matrix of a mammalian liver, liver lobe or portion thereof, comprising:
 selecting a perfusion decellularized extracellular matrix of a mammalian liver, a liver lobe or a portion thereof and two populations of cells including a first population of mammalian cells including endothelial cells or stem or progenitor cells capable of differentiation into endothelial cells, and a second population of mammalian cells including islet cells, beta cells, insulin like cells, or stem cells or progenitor cells capable of differentiation into islet cells or beta cells, wherein the selected perfusion decellularized extracellular matrix is a portion >8 cm 3  of a perfusion decellularized extracellular matrix of a liver lobe; and   contacting the perfusion decellularized extracellular matrix and the two population of cells under conditions and for a period of time that provide for re endothelialization of the vasculature of the perfusion decellularized extracellular matrix and recellularization of the perfusion decellularized extracellular matrix with the islet cells, beta cells or insulin like cells, or recellularization and differentiation and functional maturation of the stem or progenitor cells into islet cells, beta cells or insulin like cells in the perfusion decellularized extracellular matrix.   
     
     
         2 . The method of  claim 1  wherein the selected perfusion decellularized extracellular matrix is a perfusion decellularized extracellular matrix of a liver. 
     
     
         3 . The method of  claim 1  wherein the selected perfusion decellularized extracellular matrix is a perfusion decellularized extracellular matrix of a liver lobe. 
     
     
         4 . (canceled) 
     
     
         5 . The method of  claim 1  wherein the cells in the first and the second populations are xenogeneic to the decellularized extracellular matrix. 
     
     
         6 . The method of  claim 1  wherein the cells in the first and the second populations are allogeneic to the decellularized extracellular matrix. 
     
     
         7 . The method of  claim 1  wherein the cells in the first population, the second population, or both, include iPS cells. 
     
     
         8 . The method of  claim 1  wherein the perfusion decellularized extracellular matrix contains an intact vascular network. 
     
     
         9 . The method of  claim 1  wherein the conditions include perfusing the matrix with media. 
     
     
         10 . The method of  claim 9  wherein the media contain activators or inhibitors of differentiation pathways selected to provide for cell-specific differentiation including Activin A, Retinoic Acid, bFGF, clozapine-N-oxide, phosphoinositide-3-kinase inhibitor, Nicotinamide, or a combination thereof. 
     
     
         11 . The method of  claim 1  wherein the first population of cells is contacted with the extracellular matrix either by injection or perfusion, or a combination thereof. 
     
     
         12 - 14 . (canceled) 
     
     
         15 . The method of  claim 1  wherein the first population, the second population, or both, comprise human embryonic stem cells. 
     
     
         16 . The method of  claim 1  wherein the insulin producing cells are encapsulated. 
     
     
         17 . The method of  claim 1  wherein the extracellular matrix is from a nonhuman mammal and is populated with human cells. 
     
     
         18 . The method of  claim 1  further comprising implanting the extracellular matrix in a mammal. 
     
     
         19 . The method of  claim 18  wherein the extracellular matrix is implanted in the mammal prior to contact with the first population, the second population, or both. 
     
     
         20 . The method of  claim 19  wherein the cells in the first population, the second population, or both, are injected into the mammal. 
     
     
         21 . The method of  claim 18  wherein the extracellular matrix is implanted in a mammal after contact with the first population, but before the second population. 
     
     
         22 . The method of  claim 17  wherein extracellular matrix is implanted in a mammal after contact with the second population but before contact with the first population. 
     
     
         23 . The method of  claim 17  wherein the extracellular matrix is implanted in a mammal after contact with the first population and the second population. 
     
     
         24 . (canceled) 
     
     
         25 . A method to enhance insulin control in a mammal that lacks or has reduced insulin control, comprising:
 providing a re-endothelialized extracellular matrix of a mammalian liver, a liver lobe or a portion thereof having a population of mammalian cells including islet cells, beta cells, islet like cells, or stem cells or progenitor cells capable of differentiation into islet cells or beta cells, prepared by introducing mammalian endothelial cells or cells capable of differentiation into mammalian endothelial cells and the population to a decellularized extracellular matrix of the mammalian liver, liver lobe or portion thereof;   and   introducing to a mammal that lacks or has reduced insulin control the re endothelialized matrix having the population of cells including islet cells, beta cells, islet like cells or stem cells or progenitor cells capable of differentiation into islet or beta cells, so as to provide for control of blood glucose in the mammal; or   providing a re-endothelialized extracellular matrix of a mammalian liver, a liver lobe or a portion thereof prepared by introducing mammalian endothelial cells or cells capable of differentiation into mammalian endothelial cells, to a decellularized extracellular matrix of mammalian liver, liver lobe or portion thereof;   introducing to a mammal that lacks or has reduced insulin control the re endothelialized matrix; and   introducing to the mammal having the re-endothelialized matrix a population of cells including islet cells, beta cells, islet like cells or stem cells   or   progenitor cells capable of differentiation into islet or beta cells, in an amount effective to provide for control of blood glucose in the mammal; or   providing a mammal that lacks or has reduced insulin control and has an implanted re-endothelialized matrix of a mammalian liver, a liver lobe or a portion thereof, prepared by introducing mammalian endothelial cells to a decellularized extracellular matrix of a mammalian liver, liver lobe or portion thereof; and   introducing to the mammal a population of cells including islet cells, beta cells, islet like cells, or stem cells or progenitor cells capable of differentiation into islet cells, in an amount effective to provide for control of blood glucose in the mammal.   
     
     
         26 - 28 . (canceled)