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 . A pharmaceutical composition comprising a plurality of human cells augmented with isolated human mitochondria, wherein the isolated human mitochondria have undergone at least one freeze-thaw cycle prior to augmentation of the cells, and wherein the augmented human cells:
 (a) have at least 50% higher mitochondrial content compared to corresponding cells that have not been augmented and/or as compared to corresponding cells that have been augmented with mitochondria that have not undergone a freeze-thaw cycle;   (b) have an above-normal content or activity level of citrate synthase compared to corresponding cells augmented with mitochondria that have not undergone a freeze-thaw cycle;   (c) have an above-normal rate of oxygen (O 2 ) consumption compared to corresponding cells augmented with mitochondria that have not undergone a freeze-thaw cycle;   (d) have a higher rate of adenosine triphosphate (ATP) production compared to corresponding cells augmented with mitochondria that have not undergone a freeze-thaw cycle; or   (e) any combination of (a)-(d). (Original) The pharmaceutical composition of  claim 1 , wherein the human cells are bone marrow cells.   
     
     
         3 . The pharmaceutical composition of claim  2 , wherein the bone marrow cells are CD34+ cells. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein the human cells are isolated from a subject with a mitochondrial disease or disorder. 
     
     
         5 . The pharmaceutical composition of  claim 1 , wherein the isolated mitochondria are obtained from a human tissue selected from the group consisting of placenta, placental cells grown in culture, and blood cells. 
     
     
         6 . A pharmaceutical composition comprising a plurality of bone marrow cells augmented with isolated human mitochondria, wherein the isolated human mitochondria have undergone at least one freeze-thaw cycle and wherein the plurality of bone marrow cells comprise the isolated human mitochondria within the cells. 
     
     
         7 . The pharmaceutical composition of  claim 6 , wherein the bone marrow cells:
 (a) have at least 50% higher mitochondrial content compared to corresponding cells that have not been augmented and/or as compared to corresponding cells that have been augmented with mitochondria that have not undergone a freeze-thaw cycle;   (b) have an above-normal content or activity level of citrate synthase compared to corresponding cells augmented with mitochondria that have not undergone a freeze-thaw cycle;   (c) have an above-normal rate of oxygen (O 2 ) consumption compared to corresponding cells augmented with mitochondria that have not undergone a freeze-thaw cycle;   (d) have a higher rate of adenosine triphosphate (ATP) production compared to corresponding cells augmented with mitochondria that have not undergone a freeze-thaw cycle; or   (e) any combination of (a)-(d).   
     
     
         8 . The pharmaceutical composition of  claim 6 , wherein the bone marrow cells are CD34+ cells. 
     
     
         9 . The pharmaceutical composition of  claim 6 , wherein the human cells are isolated from a subject suffering from a mitochondrial disease or disorder. 
     
     
         10 . The pharmaceutical composition of  claim 6 , wherein the isolated mitochondria are obtained from a human tissue selected from the group consisting of placenta, placental cells grown in culture, and blood cells. 
     
     
         11 . A method of treating a subject suffering from a mitochondrial disease comprising:
 administering to the subject a pharmaceutical composition of  claim 1 .   
     
     
         12 . The method of  claim 11 , wherein the mitochondrial disease or disorder is selected from the group consisting of LHON, MELAS, Pearson syndrome, Leigh syndrome, NARP, MERRF, KSS, MNGIE, Friedreich's ataxia, and Alpers disease. 
     
     
         13 . The method of  claim 12 , wherein the mitochondrial disease or disorder is Pearson syndrome, LHON or MELAS. 
     
     
         14 . The method of  claim 11 , wherein the human cells are bone marrow cells. 
     
     
         15 . The method of  claim 14 , wherein the bone marrow cells are CD34+ cells. 
     
     
         16 . The method of  claim 11 , wherein the human cells are autologous. 
     
     
         17 . The method of  claim 11 , wherein the isolated mitochondria are obtained from a human tissue selected from the group consisting of placenta, placental cells grown in culture, and blood cells. 
     
     
         18 . A method of treating a subject suffering from a mitochondrial disease comprising:
 administering to the subject a pharmaceutical composition of  claim 6 .   
     
     
         19 . The method of  claim 18 , wherein the mitochondrial disease or disorder is selected from the group consisting of LHON, MELAS, Pearson syndrome, Leigh syndrome, NARP, MERRF, KSS, MNGIE, Friedreich's ataxia and Alpers disease. 
     
     
         20 . The method of  claim 19 , wherein the mitochondrial disease or disorder is Pearson syndrome, LHON or MELAS. 
     
     
         21 . The method of  claim 18 , wherein the bone marrow cells are CD34+ cells. 
     
     
         22 . The method of  claim 18 , wherein the bone marrow cells are autologous. 
     
     
         23 . The method of  claim 18 , wherein the isolated mitochondria are obtained from a human tissue selected from the group consisting of placenta, placental cells grown in culture, and blood cells. 
     
     
         24 . A pharmaceutical composition comprising a plurality of human bone marrow cells augmented with isolated human mitochondria, wherein the isolated human mitochondria have undergone at least one freeze-thaw cycle prior to augmentation of the cells, and wherein the augmented human bone marrow cells have at least 50% lower heteroplasmy level and/or at least 50% higher mitochondrial content as compared to a composition of corresponding cells that have not been augmented and/or as compared to a composition of corresponding cells that have been augmented with mitochondria that have not undergone a freeze-thaw cycle. 
     
     
         25 . The pharmaceutical composition of  claim 24 , wherein the bone marrow cells are CD34+ cells. 
     
     
         26 . The pharmaceutical composition of  claim 24 , wherein the human cells are isolated from a subject suffering from a mitochondrial disease or disorder. 
     
     
         27 . The pharmaceutical composition of  claim 24 , wherein the isolated mitochondria are obtained from a human tissue selected from the group consisting of placenta, placental cells grown in culture, and blood cells. 
     
     
         28 . The pharmaceutical composition of  claim 1 , wherein the human cells are stem cells. 
     
     
         29 . The pharmaceutical composition of  claim 24 , wherein the human cells are stem cells. 
     
     
         30 . A pharmaceutical composition comprising a plurality of human cells augmented with isolated healthy human mitochondria, wherein the augmented human cells are bone marrow or fibroblast cells, and wherein optionally the isolated healthy human mitochondria have undergone at least one freeze-thaw cycle prior to augmentation of the cells. 
     
     
         31 . The pharmaceutical composition of  claim 30 , wherein the human cells are bone marrow cells. 
     
     
         32 . The pharmaceutical composition of  claim 31 , wherein the bone marrow cells are CD34+ cells. 
     
     
         33 . The pharmaceutical composition of  claim 30 , wherein the human cells are isolated from a subject with a mitochondrial disease or disorder. 
     
     
         34 . The pharmaceutical composition of  claim 30 , wherein the isolated mitochondria are obtained from a human tissue selected from the group consisting of placenta, placental cells grown in culture, and blood cells. 
     
     
         35 . A method of treating a subject suffering from a mitochondrial disease comprising: administering to the subject a pharmaceutical composition of  claim 30 . 
     
     
         36 . The method of  claim 35 , wherein the mitochondrial disease or disorder is selected from the group consisting of LHON, MELAS, Pearson syndrome, Leigh syndrome, NARP, MERRF, KSS, MNGIE, Friedreich's ataxia, and Alpers disease. 
     
     
         37 . The method of  claim 36 , wherein the mitochondrial disease or disorder is Pearson syndrome, LHON or MELAS. 
     
     
         38 . The method of  claim 35 , wherein the human cells are bone marrow cells. 
     
     
         39 . The method of  claim 38 , wherein the bone marrow cells are CD34+ cells. 
     
     
         40 . The method of  claim 38 , wherein the human cells are autologous. 
     
     
         41 . The method of  claim 35 , wherein the isolated mitochondria are obtained from a human tissue selected from the group consisting of placenta, placental cells grown in culture, and blood cells. 
     
     
         42 . A pharmaceutical composition comprising a plurality of human cells augmented with isolated healthy human mitochondria from placenta cells. 
     
     
         43 . The pharmaceutical composition of  claim 42 , wherein the human cells are bone marrow cells. 
     
     
         44 . The pharmaceutical composition of  claim 43 , wherein the bone marrow cells are CD34+ cells. 
     
     
         45 . The pharmaceutical composition of  claim 42 , wherein the human cells are isolated from a subject with a mitochondrial disease or disorder. 
     
     
         46 . The pharmaceutical composition of  claim 42 , wherein the augmented human cells are bone marrow or fibroblast cells, and wherein optionally the isolated healthy human mitochondria from placenta cells have undergone at least one freeze-thaw cycle prior to augmentation of the cells. 
     
     
         47 . A method of treating a subject suffering from a mitochondrial disease comprising: administering to the subject a pharmaceutical composition of  claim 42 .