Mammalian cells enriched with functional mitochondria

Abstract

The present invention provides human bone-marrow cells enriched with functional mitochondria, methods for their production, and therapeutic methods utilizing such cells.

1 . An ex-vivo method for enriching human bone-marrow cells with functional mitochondria, the method comprising the steps of:
 (i) providing a first composition, comprising a plurality of human bone-marrow cells obtained or derived from a patient afflicted with a mitochondrial disease or from a subject not afflicted with a mitochondrial disease;   (ii) providing a second composition, comprising a plurality of isolated human functional mitochondria obtained from a subject not afflicted with a mitochondrial disease;   (iii) contacting the human bone-marrow cells of the first composition with the human functional mitochondria of the second composition, thus forming a third composition; and   (iv) incubating the third composition under conditions allowing the human functional mitochondria to enter the human bone-marrow cells thereby enriching said human bone-marrow cells with said human functional mitochondria, thus forming a fourth composition;   wherein the mitochondrial content of the human bone-marrow cells in the fourth composition is at least 50% higher than the mitochondrial content of the human bone-marrow cells in the first composition.   
     
     
         2 . The method of  claim 1 , wherein the mitochondrial content of the bone-marrow cells in the first composition or in the fourth composition is determined by determining the content or activity level of citrate synthase. 
     
     
         3 - 8 . (canceled) 
     
     
         9 . The method of  claim 1 , wherein the bone-marrow cells express the bone-marrow progenitor cell antigen CD34 (CD34+). 
     
     
         10 - 11 . (canceled) 
     
     
         12 . The method of  claim 1 , further comprising concentrating the bone-marrow cells and the functional mitochondria in the third composition before or during incubation. 
     
     
         13 . The method of  claim 12 , further comprising centrifugation of the third composition before, during or after incubation. 
     
     
         14 . The method of  claim 1 , wherein the bone-marrow cells in the first composition are obtained from a patient afflicted with a mitochondrial disease, and have:
 (i) a sub-normal rate of oxygen (O2) consumption;   (ii) a sub-normal content or activity level of citrate synthase;   (iii) a sub-normal rate of adenosine triphosphate (ATP) production; or   (iv) any combination of (i), (ii) and (iii).   
     
     
         15 . (canceled) 
     
     
         16 . The method of  claim 1 , wherein the bone-marrow cells in the first composition are obtained from a subject not afflicted with a mitochondrial disease, and have:
 (i) a normal rate of oxygen (O2) consumption;   (ii) a normal content or activity level of citrate synthase;   (iii) a normal rate of adenosine triphosphate (ATP) production; or   (iv) any combination of (i), (ii) and (iii).   
     
     
         17 . The method of  claim 1 , wherein the isolated human functional mitochondria in the second composition are obtained from a subject not afflicted with a mitochondrial disease, and have:
 (i) a normal rate of oxygen (O2) consumption;   (ii) a normal content or activity level of citrate synthase;   (iii) a normal rate of adenosine triphosphate (ATP) production; or   (iv) any combination of (i), (ii) and (iii).   
     
     
         18 . The method of  claim 1 , wherein the bone-marrow cells in the fourth composition have:
 (i) an above-normal rate of oxygen (O2) consumption;   (ii) an above-normal content or activity level of citrate synthase;   (iii) an above-normal rate of adenosine triphosphate (ATP) production; or   (iv) any combination of (i), (ii) and (iii).   
     
     
         19 . (canceled) 
     
     
         20 . The method of  claim 1 , wherein the fourth composition is not enriched with cytochrome C reductase or cytochrome C reductase activity compared to the first composition. 
     
     
         21 - 25 . (canceled) 
     
     
         26 . A plurality of human bone-marrow cells enriched with functional mitochondria, obtained by the method of  claim 1 . 
     
     
         27 . A plurality of human bone-marrow cells, wherein the bone-marrow cells:
 (a) have an above-normal mitochondrial content;   (b) have an above-normal rate of oxygen (O2) consumption;   (c) have an above-normal content or activity level of citrate synthase;   (d) are CD34+; or   (e) any combination of (a), (b), (c) and (d).   
     
     
         28 . The plurality of human bone-marrow cells of  claim 27 , having an above-normal mitochondrial content; having an above-normal rate of oxygen (O2) consumption; having an above-normal content or activity level of citrate synthase; and are CD34+. 
     
     
         29 . A pharmaceutical composition comprising a plurality of human bone-marrow cells according to  claim 26 . 
     
     
         30 . (canceled) 
     
     
         31 . A method of treating a mitochondrial disease in a human patient in need thereof, comprising the step of administering to the patient the pharmaceutical composition of  claim 29 . 
     
     
         32 - 38 . (canceled) 
     
     
         39 . The method of  claim 31 , wherein the mitochondrial disease is associated with a mutation in the mitochondrial DNA. 
     
     
         40 - 43 . (canceled) 
     
     
         44 . The method of  claim 31 , wherein the mitochondrial disease is LHON. 
     
     
         45 . The method of  claim 31 , wherein the mitochondrial disease is MELAS.