Methods of controlling proliferation and differentiation of stem and progenitor cells

Abstract

A method of expanding a population of cells, while at the same time inhibiting differentiation of the cells, the method includes the step of providing the cells with conditions for cell proliferation and, at the same time, for reducing a capacity of the cells in utilizing copper. The method can be executed both in-vivo and ex-vivo. A method of inducing differentiation in a population of cells, the method includes the step of providing the cells with a transition metal chelator which binds copper and which is effective in inducing cell differentiation.

What is claimed is:  
     
         1 . A method of hematopoietic cells transplantation comprising the steps of: 
 (a) obtaining hematopoietic cells to be transplanted from a donor;    (b) providing said cells ex-vivo with conditions for cell proliferation and, at the same time, for reducing a capacity of said cells in utilizing cooper, thereby expanding a population of said cells, while at the same time, inhibiting differentiation of said cells; and    (c) transplanting said cells to a patient.    
     
     
         2 . The method of  claim 1 , wherein said donor and said patient are a single individual.  
     
     
         3 . The method of  claim 1 , wherein obtaining said hematopoetic cells is from a source selected from the group consisting of peripheral blood, bone marrow, neonatal umbilical cord blood and embryonic stem cells.  
     
     
         4 . The method of  claim 3 , wherein obtaining said hematopoietic cells further includes enriching said cells for stem cells.  
     
     
         5 . The method of  claim 3 , wherein obtaining said hematopoietic cells further includes enriching said cells for progenitor cells.  
     
     
         6 . The method of  claim 1 , wherein reducing said capacity of the cells in utilizing copper is effected by a transition metal chelator having an affinity to copper.  
     
     
         7 . The method of  claim 6 , wherein said transition metal chelator is selected from the group consisting of polyamine chelating agents, ethylendiamine, diethylenetriamine, triethylenetetramine, triethylenediamine, tetraethylenepentamine, aminoethylethanolamine, aminoethylpiperazine, pentaethylenehexamine, t triethylenetetramine-hydrochloride, tetraethylenepentamine-hydrochloride, pentaethylenehexamine-hydrochloride, tetraethylpentamine, captopril, penicilamine, N,N′-bis(3-aminopropyl)-1,3-propanediamine, N,N,Bis (2 animoethyl) 1,3 propane diamine, 1,7-dioxa-4,10-diazacyclododecane, 1,4,8,11-tetraaza cyclotetradecane-5,7-dione, 1,4,7-triazacyclononane trihydrochloride, 1-oxa-4,7,10-triazacyclododecane, 1,4,8,12-tetraaza cyclopentadecane, 1,4,7,10-tetraaza cyclododecane.  
     
     
         8 . The method of  claim 1 , wherein providing the cells with said conditions for cell proliferation include providing the cells with nutrients and with cytokines.  
     
     
         9 . The method of  claim 8 , wherein said cytokines are early acting cytokines.  
     
     
         10 . The method of  claim 9 , wherein said early acting cytokines are selected from the group consisting of stem cell factor, FLT3 ligand, interleukin-6, thrombopoietin and interleukin-3.  
     
     
         11 . The method of  claim 8 , wherein said cytokines are late acting cytokines.  
     
     
         12 . The method of  claim 11 , wherein said late acting cytokines are selected from the group consisting of granulocyte colony stimulating factor, granulocyte/macrophage colony stimulating factor and erythropoietin.  
     
     
         13 . The method of  claim 1 , wherein said cells are derived from a source selected from the group consisting of bone marrow, peripheral blood and neonatal umbilical cord blood.  
     
     
         14 . The method of  claim 1 , wherein said cells are enriched for hematopoietic CD 34 + cells.  
     
     
         15 . The method of  claim 1 , wherein said cells are selected from the group consisting of non-differentiated stem cells and committed progenitor cells.  
     
     
         16 . A method of genetically modifying stem cells with an exogene comprising the steps of: 
 (a) obtaining stem cells to be genetically modified;    (b) providing said cells ex-vivo with conditions for cell proliferation and, at the same time, for reducing a capacity of said cells in utilizing cooper, thereby expanding a population of said cells, while at the same time, inhibiting differentiation of said cells; and    (c) genetically modifying said cells with the exogene.    
     
     
         17 . The method of  claim 16 , wherein genetically modifying is effected by a vector including the exogene.  
     
     
         18 . The method of  claim 16 , wherein reducing said capacity of the cells in utilizing copper is effected by a transition metal chelator having an affinity to copper.  
     
     
         19 . The method of  claim 18 , wherein said transition metal chelator is selected from the group consisting of polyamine chelating agents, ethylendiamine, diethylenetriamine, triethylenetetramine, triethylenediamine, tetraethylenepentamine, aminoethylethanolamine, aminoethylpiperazine, pentaethylenehexamine, tethylenetetramine-hydrochloride, tetraethylenepentamine-hydrochloride, pentaethylenehexamine-hydrochloride, tetraethylpentamine, captopril, penicilamine, N,N′-bis(3-aminopropyl)-1,3-propanediamine, N,NBis (2 animoethyl) 1,3 propane diamine, 1,7-dioxa-4,10-diazacyclododecane, 1,4,8,11-tetraaza cyclotetradecane-5,7-dione, 1,4,7-triazacyclononane trihydrochloride, 1-oxa-4,7,10-triazacyclododecane, 1,4,8,12-tetraaza cyclopentadecane, 1,4,7,10-tetraaza cyclododecane.  
     
     
         20 . The method of  claim 16 , wherein providing the cells with said conditions for cell proliferation include providing the cells with nutrients and with cytokines.  
     
     
         21 . The method of  claim 20 , wherein said cytokines are early acting cytokines.  
     
     
         22 . The method of  claim 21 , wherein said early acting cytokines are selected from the group consisting of stem cell factor, FLT3 ligand, interleukin-6, thrombopoietin and interleukin-3.  
     
     
         23 . The method of  claim 20 , wherein said cytokines are late acting cytokines.  
     
     
         24 . The method of  claim 23 , wherein said late acting cytokines are selected from the group consisting of granulocyte colony stimulating factor, granulocyte/macrophage colony stimulating factor and erythropoietin.  
     
     
         25 . The method of  claim 16 , wherein said cells are derived from a source selected from the group consisting of bone marrow, peripheral blood and neonatal umbilical cord blood.  
     
     
         26 . A method of adoptive immunotherapy comprising the steps of: 
 (a) obtaining progenitor hematopoietic cells from a patient;    (b) providing said cells ex-vivo with conditions for cell proliferation and, at the same time, for reducing a capacity of said cells in utilizing cooper, thereby expanding a population of said cells, while at the same time, inhibiting differentiation of said cells; and    (c) transplanting said cells to the patient.    
     
     
         27 . The method of  claim 26 , wherein reducing said capacity of the cells in utilizing copper is effected by a transition metal chelator having an affinity to copper.  
     
     
         28 . The method of  claim 27 , wherein said transition metal chelator is selected from the group consisting of polyamine chelating agents, ethylendiamine, diethylenetriamine, triethylenetetramine, triethylenediamine, tetraethylenepentamine, aminoethylethanolamine, aminoethylpiperazine, pentaethylenehexamine, triethylenetetramine-hydrochloride, tetraethylenepentamine-hydrochloride, pentaethylenehexamine-hydrochloride, tetraethylpentamine, captopril, penicilamime, N,N′-bis(3-aminopropyli 1,3-propanediamine, N,N,Bis (2 animoethyl) 1,3 propane diamine, 1,7-dioxa-4,10-diazacyclododecane, 1,4,8,11-tetraaza cyclotetradecane-5,7-dione, 1,4,7-triazacyclononane trihydrochloride, 1-oxa-4,7,10-triazacyclododecane, 1,4,8,12-tetraaza cyclopentadecane, 1,4,7,10-tetraaza cyclododecane.  
     
     
         29 . The method of  claim 26 , wherein providing the cells with said conditions for cell proliferation include providing the cells with nutrients and with cytokines.  
     
     
         30 . The method of  claim 29 , wherein said cytokines are early acting cytokines.  
     
     
         31 . The method of  claim 30 , wherein said early acting cytokines are selected from the group consisting of stem cell factor, FLT3 ligand, interleukin-6, thrombopoietin and interleukin-3.  
     
     
         32 . The method of  claim 29 , wherein said cytokines are late acting cytokines.  
     
     
         33 . The method of  claim 32 , wherein said late acting cytokines are selected from the group consisting of granulocyte colony stimulating factor, granulocyte/macrophage colony stimulating factor and erythropoietin.  
     
     
         34 . The method of  claim 26 , wherein said cells are derived from a source selected from the group consisting of bone marrow, peripheral blood and neonatal umbilical cord blood.  
     
     
         35 . The method of  claim 26 , wherein said cells are enriched for hematopoietic CD 34 + cells.  
     
     
         36 . The method of  claim 26 , wherein said cells are selected from the group consisting of non-differentiated stem cells and committed progenitor cells.