Preparative regimen for engraftment, growth and differentiation of non-hematopoeitic cells in vivo after transplantation

Abstract

The invention relates to methods of obtaining an expanded population of mammalian ex vivo cells and/or for treating a mammalian subject by (a) administering to a subject an effective amount of an agent that confers a growth disadvantage to at least a subset of endogenous cells at the site of engraftment; (b) administering to the subject an effective amount of a mitogenic stimulus for the ex vivo cells; and (c) administering the ex vivo cells to the subject, wherein the ex vivo cells engraft at the site and proliferate to a greater extent than the subset of endogenous cells, to repopulate at least a portion of the engraftment site with the ex vivo cells. The repopulated cells can be harvested for further use or be left at the engraftment site of a subject to be treated. The invention also provides methods of treating brain injury in a subject by engrafting ex vivo cells at the site of injury.

1 . A method of treating a mammalian subject with an organ having a reduced function by grafting mammalian ex vivo cells which supplement the function and by promoting proliferation of the engrafted cells at the site of engraftment, the method comprising the steps of:
 (a) administering to the subject an effective amount of an agent that confers a growth disadvantage to at least a subset of endogenous cells at the site of engraftment;   (b) administering to the subject an effective amount of a mitogenic stimulus for the ex vivo cells; and   (c) administering the ex vivo cells to the subject, wherein the ex vivo cells engraft at the site and proliferate to a greater extent than the subset of endogenous cells, to repopulate at least a portion of the engraftment site with the ex vivo cells, wherein the repopulated ex vivo cells supplement or provide the function.   
     
     
         2 . The method of  claim 1 , wherein the organ with the reduced function is the intestine, liver, lung, kidney, pancreas, eye, testes, ovary, brain, spinal cord, or skin. 
     
     
         3 . The method of  claim 1 , wherein the organ with the reduced function is selected from the group consisting of intestine and brain. 
     
     
         4 . The method of  claim 1 , wherein the organ with the reduced function was damaged by irradiation, trauma, chemical or drug exposure, a genetic illness, or an infectious disease. 
     
     
         5 . The method of  claim 1 , wherein the ex vivo cells are selected from the group consisting of adult somatic cells, adult progenitor cells, adult stem cells, embryonic progenitor cells, fetal cells, and embryonic stem cells from the same species as the subject or from a different mammalian species than the subject. 
     
     
         6 . The method of  claim 1 , wherein the ex vivo cells are heterologous to the subject. 
     
     
         7 . The method of  claim 1 , wherein the ex vivo cells are autologous to the subject. 
     
     
         8 . The method of  claim 1 , wherein the ex vivo cells are cultured, expanded or modified prior to their administration to the subject. 
     
     
         9 . The method of  claim 1 , wherein the ex vivo cells are harvested and sorted from a heterogeneous cell population prior to their administration to the subject. 
     
     
         10 . The method of  claim 1 , wherein the agent that confers a growth disadvantage to at least a subset of endogenous cells is radiation. 
     
     
         11 . The method of  claim 10 , wherein the radiation is selected from the group consisting of x-ray and gamma ray. 
     
     
         12 . The method of  claim 1 , wherein the agent that confers a growth disadvantage to at least a subset of endogenous cells is selected from the group consisting of cytotoxic chemicals, ultrasound, heat, biological agents, misfolded proteins, and proteins or nucleic acids that suppress cell division. 
     
     
         13 . The method of  claim 1 , wherein the protein that suppresses cell division is an antibody. 
     
     
         14 . The method of  claim 1 , wherein the mitogenic stimuli comprises one or more growth factors selected from the group consisting of HGF, EGF, FGF, VEGF, NGF, 1′-6, TNF-alpha, CTNF, R-spondin 1, Noggin, and TWEAK. 
     
     
         15 - 18 . (canceled) 
     
     
         19 . The method of  claim 1 , wherein the organ with reduced function was damaged by a neurodegenerative disease. 
     
     
         20 - 31 . (canceled) 
     
     
         32 . A method of obtaining an expanded population of ex vivo cells, comprising the steps of:
 (a) administering to a non-human mammalian subject an effective amount of an agent that confers a growth disadvantage to at least a subset of endogenous cells at a site of engraftment;   (b) administering to the non-human mammalian subject an effective amount of a mitogenic stimuli for the ex vivo cells; and   (c) administering the ex vivo cells t the subject, wherein the ex vivo cells engraft at the site and proliferate to a greater extent than the subset of endogenous cells, thereby repopulating at least a portion of the engraftment site with the ex vivo cells, and   (d) harvesting the repopulated cells from the engraftment site;   
       thereby obtaining the expanded ex vivo cell population. 
     
     
         33 . The method of  claim 32 , wherein the ex vivo cells are human. 
     
     
         34 . The method of  claim 32 , wherein the ex vivo cells are selected from the group consisting of adult somatic cells, adult progenitor cells, adult stem cells, embryonic progenitor cells, fetal cells, xenogenic cells, and embryonic stem cells. 
     
     
         35 . The method of  claim 32 , wherein the ex vivo cells heterologous to the non-human subject. 
     
     
         36 . The method of  claim 32 , wherein the agent that confers a growth disadvantage to at least a subset of endogenous cells of the organ is radiation. 
     
     
         37 . The method of  claim 32 , wherein the radiation is selected from the group consisting of x-ray and gamma ray. 
     
     
         38 . The method of  claim 32 , wherein the agent that confers a growth disadvantage to at least a subset of endogenous cells of the damaged organ is selected from the group consisting of cytotoxic chemicals, ultrasound, heat, biological agents, degenerative proteins, and proteins that suppress cell division. 
     
     
         39 . The method of  claim 32 , wherein the mitogenic composition comprises a growth factor selected from the group consisting of HGF, EGF, FGF, VEGF, NGF, 11-6, TNF-alpha, CTNF, R-spondin 1, Noggin, and TWEAK. 
     
     
         40 . The method of  claim 32 , wherein the ex vivo cell is derived from brain, intestinal, or liver tissue. 
     
     
         41 . A method of treating a subject having an organ with reduced function, comprising engrafting cells obtained by the method of  claim 32 . 
     
     
         42 . A method of treating a subject having an organ with reduced function, comprising engrafting ex vivo cells obtained by the method of  claim 33 . 
     
     
         43 . The method of  claim 42 , wherein the organ is intestine, liver, lung, kidney, pancreas, eye, testes, brain, ovary, or skin. 
     
     
         44 . A method of treating a mammalian subject having a damaged central nervous system with a reduced function by engrafting mammalian ex vivo cells at the site of injury, the method comprising the steps of:
 (a) administering to the subject an effective amount of an agent that increases the engraftment of ex vivo cells at the site of injury;   (b) optionally administering to the subject an effective amount of a mitogenic stimuli for the ex vivo cells; and   (c) administering the ex vivo cells to the subject, wherein the ex vivo cells engraft at the site of injury and repopulate at least a portion of the site with the ex vivo cells, wherein the repopulated ex vivo cells supplement the function,   thereby treating the subject.   
     
     
         45 . The method of  claim 44 , wherein the ex vivo cells are selected from the group consisting of adult somatic cells, adult neuron progenitor cells, adult stem cells, embryonic progenitor cells, fetal cells, xenogenic cells, and embryonic stem cells which are capable of populating the site of injury with neurons. 
     
     
         46 . A method of obtaining an expanded population of ex vivo cells, comprising the steps of:
 (a) administering to an irradiated organ ex vivo or in culture an effective amount of a mitogenic stimuli for the ex vivo cells; wherein the irradiation confers a growth disadvantage to at least a subset of endogenous cells at a site where the ex vivo cells engraft; and   (c) administering the ex vivo cells to the irradiated organ, wherein the ex vivo cells engraft at the site and proliferate to a greater extent than the subset of endogenous cells, thereby repopulating at least a portion of the engraftment site with the ex vivo cells, and   (d) harvesting the repopulated cells from the engraftment site;   
       thereby obtaining the expanded ex vivo cell population. 
     
     
         47 . The method of  claim 46 , wherein the ex vivo cells are human. 
     
     
         48 . The method of  claim 46 , wherein the ex vivo cells are selected from the group consisting of adult somatic cells, adult progenitor cells, adult stem cells, embryonic progenitor cells, fetal cells, xenogenic cells, and embryonic stem cells. 
     
     
         49 . The method of  claim 46 , wherein the ex vivo cell is heterologous to the non-human subject. 
     
     
         50 . The method of  claim 46 , wherein the agent that confers a growth disadvantage to at least a subset of endogenous cells of the organ is radiation. 
     
     
         51 . The method of  claim 46 , wherein the radiation is selected from the group consisting of x-ray and gamma ray. 
     
     
         52 . The method of  claim 46 , wherein the agent that confers a growth disadvantage to at least a subset of endogenous cells of the damaged organ is selected from the group consisting of cytotoxic chemicals, ultrasound, heat, biological agents, degenerative proteins, and proteins that suppress cell division. 
     
     
         53 . The method of  claim 46 , wherein the mitogenic composition comprises a growth factor selected from the group consisting of HGF, EGF, FGF, VEGF, NGF, 11-6, TNF-alpha, CTNF, R-spondin 1, Noggin, and TWEAK. 
     
     
         54 . The method of  claim 46 , wherein the ex vivo cell is derived from brain, intestinal, or liver tissue.