Target populations of oligodendrocyte precursor cells and methods of making and using same

Abstract

This application provides for enriched target populations oligodendrocyte precursor cells (OPCs) that can differentiate into oligodendrocytes. The target OPCs may be expanded and optionally subjected to conditions to induce their differentiation into oligodendrocytes. The target OPCs and their progeny are useful for the treatment of disease associated with demyelination of central nervous system axons.

1 . An enriched population of oligodendrocyte precursor cells (OPCs) derived from neural or neural derived cells, wherein the population is enriched for target OPCs that are PDGFRα +  and CD105 − , wherein at least 30% of the cells in the population are target OPCs. 
   
   
       2 . The population of  claim 1 , where in the target cells are also PSA-NCAM lo/− . 
   
   
       3 . The population of  claim 1 , where in the target cells are also A2B5 lo/− . 
   
   
       4 . The population of  claim 1 , where in the target cells are also CD133 + . 
   
   
       5 . The population of OPCs in  claim 1 , wherein at least 50% of the cells in the population are target OPCs. 
   
   
       6 . The population of OPCs in  claim 1 , wherein at least 70% of the cells in the population are target OPCs. 
   
   
       7 . The population of OPCs in  claim 1 , wherein at least 90% of the cells in the population are target OPCs. 
   
   
       8 . The population of OPCs in  claim 1 , wherein the target OPCs are PDGFRα + , CD105 − , CD133 + , and A2B5 − . 
   
   
       9 . The population of OPCs in  claim 1 , wherein the target OPCs are PDGFRα + , CD105 − , CD133 + , A2B5 lo/− , and PSA-NCAM − . 
   
   
       10 . A method of producing population of cells enriched for target oligodendrocyte precursor cells (OPCs) from neural or neural derived cells, comprising contacting neural or neural derived cells with at least one reagent that binds to cell surface antigens expressed by the target OPCs, wherein the target OPCs are PDGFRα +  and CD105 − , wherein the population of cells is enriched to contain at least 30% target OPC. 
   
   
       11 . The method of  claim 10 , wherein the target OPCs are also PSA-NCAM lo/− . 
   
   
       12 . The method of  claim 10 , where in the target cells are also A2B5 lo/− . 
   
   
       13 . The method of  claim 10 , where in the target cells are also CD133 + . 
   
   
       14 . The method of  claim 10  further comprising negatively selecting for target OPCs by selectively removing nontarget cell populations. 
   
   
       15 . The method of  claim 10 , wherein the target OPCs are PDGFRα + , CD105 − , CD133 + , and A2B5 − . 
   
   
       16 . The method of  claim 10 , wherein the target OPCs are PDGFRα + , CD105 − , CD133 + , A2B5 lo/− , and PSA-NCAM − . 
   
   
       17 . A method of increasing the number of target oligodendrocyte precursor cells (OPCs) in a cell culture comprising culturing a population of cells comprising at least 30% target OPCs to increase the number of target OPCs in the cell culture, wherein the target OPCs are PDGFRα +  and CD105 − . 
   
   
       18 . The method of  claim 17 , wherein the target OPCs are also PSA-NCAM lo/− . 
   
   
       19 . The method of  claim 17 , where in the target cells are also A2B5 lo/− . 
   
   
       20 . The method of  claim 17 , where in the target cells are also CD133 + . 
   
   
       21 . The method of  claim 17 , wherein the target OPCs are PDGFRα + , CD105 − , CD133 + , and A2B5 − . 
   
   
       22 . The method of  claim 17 , wherein the target OPCs are PDGFRα + , CD105 − , CD133 + , A2B5 lo/− , and PSA-NCAM − . 
   
   
       23 . The method of  claim 17  further comprising contacting the OPCs with an effective amount of at least one biological agent that is capable of increasing the number of OPCs. 
   
   
       24 . The method of  claim 23  wherein the biological agent is selected from the group consisting of leukemia inhibitory factor (LIF), epidermal growth factor (EGF), basic fibroblast growth factor (FGF-2; bFGF), IGF-1, NT3, Shh, CTNF, PDGF-AA and combinations thereof. 
   
   
       25 . A method for proliferating oligodendrocyte precursor cells (OPCs) comprising: proliferating OPCs in serum-free culture medium containing one or more predetermined growth factors effective for inducing OPC proliferation, wherein: (a) the population comprises OPCs which are PDGFRα +  and CD105 − ; and (b) in the presence of differentiation-inducing conditions, the cells produce progeny cells that differentiate into oligodendrocytes. 
   
   
       26 . The method of  claim 25 , wherein the OPCs are also PSA-NCAM lo/− . 
   
   
       27 . The method of  claim 25 , where in the target cells are also A2B5 lo/− . 
   
   
       28 . The method of  claim 25 , where in the target cells are also CD133 + . 
   
   
       29 . The method of  claim 25  further comprising subjecting the OPCs to culture conditions that induce oligodendrocyte differentiation to produce differentiating and differentiated oligodendrocytes. 
   
   
       30 . A composition comprising the oligodendrocyte precursor cells (OPCs) produced by the method of  claim 25 . 
   
   
       31 . A composition comprising the differentiating or differentiated oligodendrocytes produced by the method of  claim 25 . 
   
   
       32 . A method of treating a mammalian individual suffering from a disease associated with demyelination of central nervous system axons, comprising introducing the enriched population of OPCs of  claim 1  to the mammalian individual in an amount effective to treat the disease. 
   
   
       33 . The method of  claim 32 , wherein the mammalian individual is a human. 
   
   
       34 . The method of  claim 32 , wherein the OPCs are administered to the mammalian individual by cell transplantation. 
   
   
       35 . The method of  claim 32 , wherein the disease is multiple sclerosis, Pelizaeus-Merzbacher disease, cerebral palsy, radiation induced myelination disorders, acute disseminated encephalomyelitis, transverse myelitis, demyelinating genetic disease, spinal cord injury, virus-induced demyelination, Progressive Multifocal Leucoencephalopathy, Human Lymphotrophic T-cell Virus I (HTLVI)-associated myelopathy, or nutritional metabolic disorder. 
   
   
       36 . The method of  claim 32 , wherein the disease is multiple sclerosis, Pelizaeus-Merzbacher disease, or cerebral palsy. 
   
   
       37 . A method of treating a mammalian individual suffering from a disease associated with demyelination of central nervous system axons, comprising introducing the OPCs of  claim 1  to the mammalian individual in an amount effective to treat the disease. 
   
   
       38 . The method of  claim 37 , wherein the mammalian individual is a human. 
   
   
       39 . The method of  claim 37 , wherein the OPCs are administered to the mammalian individual by cell transplantation. 
   
   
       40 . The method of  claim 37 , wherein the disease is multiple sclerosis, acute disseminated encephalomyelitis, transverse myelitis, demyelinating genetic disease, spinal cord injury, virus-induced demyelination, Progressive Multifocal Leucoencephalopathy, Human Lymphotrophic T-cell Virus I (HTLVI)-associated myelopathy, or nutritional metabolic disorder. 
   
   
       41 . A method of treating a mammalian individual suffering from a disease associated with demyelination of central nervous system axons, comprising introducing the differentiating or differentiated oligodendrocytes prepared according to the method of  claim 21  to the mammalian individual in an amount effective to treat the disease. 
   
   
       42 . The method of  claim 41 , wherein the mammalian individual is a human. 
   
   
       43 . The method of  claim 41 , wherein the differentiating or differentiated oligodendrocytes are administered to the mammalian individual by cell transplantation. 
   
   
       44 . The method of  claim 41 , wherein the disease is multiple sclerosis, acute disseminated encephalomyelitis, transverse myelitis, demyelinating genetic disease, spinal cord injury, virus-induced demyelination, Progressive Multifocal Leucoencephalopathy, Human Lymphotrophic T-cell Virus I (HTLVI)-associated myelopathy, or nutritional metabolic disorder. 
   
   
       45 . A method of screening for compounds that affect a biological function of an enriched population of target oligodendrocyte precursor cells comprising: (a) contacting an enriched population of target oligodendrocyte precursor cells obtained by the method of  claim 1  with a test compound; and (b) detecting a change in a biological function of the oligodendrocyte precursor cells. 
   
   
       46 . The method of  claim 45 , wherein said target oligodendrocyte precursor cells are PDGFRα + , CD105 − , A2B5 lo/− , PSA-NCAM lo/− , and CD133 + . 
   
   
       47 . The method of  claim 45 , wherein said target oligodendrocyte precursor cells are PDGFRα + , CD105 − , A2B5 − , and CD133 + . 
   
   
       48 . The method of  claim 45 , wherein said target oligodendrocyte precursor cells are PDGFRα + , CD105 − , A2B5 lo/− , PSA-NCAM − , and CD133 + . 
   
   
       49 . The method of  claim 45 , wherein the change occurs in at least one of the characteristics selected from the group consisting of myelination, differentiation into oligodendrocytes, proliferation rate, cell migration, viability, gene expression, protein expression, protein levels in the culturing medium, dedifferentiation, growth characteristics, and cell morphology. 
   
   
       50 . A method of producing a population enriched for oligodendrocyte precursor cells comprising:
 (a) contacting neural or neural derived cells comprising one or more multipotent central nervous system stem cell with an antibody that specifically binds to PDGFRα; and   (b) selecting said neural or neural derived cells that are PDGFRα hi , wherein the selected cells are enriched for oligodendrocyte precursor cells as compared with the neural or neural derived cells.   
   
   
       51 . The method of  claim 50 , wherein said neural or neural derived cells are obtained from a neuro sphere culture or an adherent culture. 
   
   
       52 . The method of  claim 50 , wherein the method further comprises the step of eliminating those cells that are PDGFRα lo/med . 
   
   
       53 . The method of  claim 50 , wherein the method further comprises the step of eliminating cells that are CD105 − .