Porcine neural cells and their use in treatment of neurological deficits due to neurodegenerative diseases

Abstract

Porcine neural cells and methods for using the cells to treat neurological deficits due to neurodegeneration are described. The porcine neural cells are preferably embryonic mesencephalic, embryonic striatal cells, or embryonic cortical cells. The porcine neural cells can be modified to be suitable for transplantation into a xenogeneic subject, such as a human. For example, the porcine neural cells can be modified such that an antigen (e.g., an MHC class I antigen) on the cell surface which is capable of stimulating an immune response against the cell in a xenogeneic subject is altered (e.g., by contact with an anti-MHC class I antibody, or a fragment or derivative thereof) to inhibit rejection of the cell when introduced into the subject. In one embodiment, the porcine neural cells are obtained from a pig which is essentially free from organisms or substances which are capable of transmitting infection or disease to the recipient subject. The porcine neural cells of the present invention can be used to treat neurological deficits due to neurodegeneration in the brain of a xenogeneic subject (e.g., a human with epilepsy, head trauma, stroke, amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, or Huntington's disease) by introducing the cells into the brain of the subject.

1 . A porcine striatal cell, which, in unmodified form, has at least one antigen on the cell surface which is capable of stimulating an immune response against the cell in a xenogeneic subject, wherein the antigen on the cell surface is altered to inhibit rejection of the cell when introduced into the xenogeneic subject.  
     
     
         2 . The porcine striatal cell of  claim 1 , wherein the antigen on the cell surface which is altered is an MHC class I antigen.  
     
     
         3 . The porcine striatal cell of  claim 2  , which is contacted prior to transplantation into a xenogeneic subject with at least one anti-MHC class I antibody, or fragment or derivative thereof, which binds to the MHC class I antigen on the cell surface but does not activate complement or induce lysis of the cell.  
     
     
         4 . The porcine striatal cell of  claim 3 , wherein the anti-MHC class I antibody is an anti-MHC class I F(ab′) 2  fragment.  
     
     
         5 . The porcine striatal cell of  claim 4 , wherein the anti-MHC class I F(ab′) 2  fragment is a F(ab′) 2  fragment of a monoclonal antibody PT85.  
     
     
         6 . The porcine striatal cell of  claim 1 , wherein the xenogeneic subject is a human.  
     
     
         7 . The porcine striatal cell of  claim 1 , which is an embryonic striatal cell.  
     
     
         8 . The isolated striatal cell of  claim 7 , which is obtained from a lateral ganglionic eminence of porcine striatum.  
     
     
         9 . The porcine striatal cell of  claim 8 , which is obtained from an embryonic pig between about days 30 and 40of gestation.  
     
     
         10 . An isolated porcine striatal cell having an antibody, antibody fragment or derivative, bound to its surface.  
     
     
         11 . The porcine striatal cell of  claim 10 , wherein the antibody is an anti-MHC class I antibody.  
     
     
         12 . The porcine striatal cell of  claim 11 , wherein the anti-MHC class I antibody is an anti-MHC class I F(ab′) 2  fragment.  
     
     
         13 . The porcine striatal cell of  claim 12  , wherein the MHC class I F(ab′) 2  fragment is a F(ab′) 2  fragment of a monoclonal antibody PT85.  
     
     
         14 . An isolated porcine striatal cell obtained from a lateral ganglionic eminence of an embryonic pig between about days 30 and 40 of gestation, the striatal cell having an antibody bound to an antigen on its surface.  
     
     
         15 . A composition comprising a porcine striatal cell and an antibody which binds to an antigen on the surface of the porcine striatal cell.  
     
     
         16 . The composition of  claim 15 , further comprising at least one of the agents or factors selected from the group consisting of an angiogenic factor, a neurotrophic factor, an antiapoptotic factor, an antioxidant, and an agent which inhibits nitric oxide production.  
     
     
         17 . The composition of  claim 16 , wherein the angiogenic factor is selected from the group consisting of vascular endothelial growth factor, platelet-derived growth factor, fibroblast growth factor, and epidermal growth factor.  
     
     
         18 . The composition of  claim 16 , wherein the neurotrophic factor is selected from the group consisting of glial cell line-derived growth factor, brain-derived neurotrophic factor, ciliary neurotrophic factor, midkine, insulin-like growth factor I and II, insulin, fibroblast growth factor, neurotrophin-3, neurotrophin 4/5, transforming growth factor β, FK506, and cyclosporine A  
     
     
         19 . The composition of  claim 16 , wherein the antiapoptotic factor is selected from the group consisting of the bcl-2 gene product, the bcl-xL gene product, the bcl-Xβ gene product, the crmA gene product, nerve growth factor, platelet-derived growth factor, superoxide dismutase, catalase, and N-acetyl cysteine.  
     
     
         20 . The composition of  claim 16 , wherein the antioxidant is selected from the group consisting of superoxide dismutase, glutathione peroxidase, N-acetyl cysteine, a lazaroid, vitamin C, vitamin E, and beta carotene.  
     
     
         21 . The composition of  claim 16 , wherein the agent which inhibits nitric oxide production is selected from the group consisting of gangliosides, FK506, cyclosporine A, and superoxide dismutase.  
     
     
         22 . An isolated porcine striatal cell.  
     
     
         23 . The porcine striatal cell of  claim 22 , which is obtained from- embryonic porcine striatum.  
     
     
         24 . The porcine striatal cell of  claim 23 , which is obtained from a lateral ganglionic eminence of porcine striatum.  
     
     
         25 . The porcine striatal cell of  claim 24 , which is obtained from an embryonic pig between about days 30 and 40 of gestation.  
     
     
         26 . An isolated population of porcine striatal cells.  
     
     
         27 . The population of cells of  claim 26 , wherein the cells are obtained from embryonic porcine striatum.  
     
     
         28 . The population of cells of  claim 27 , wherein the cells are obtained from a ganglionic eminence of porcine striatum.  
     
     
         29 . The population of cells of  claim 28 , wherein the cells are obtained from a lateral ganglionic eminence of porcine striatum.  
     
     
         30 . The population of cells of  claim 26 , comprising neural progenitor cells.  
     
     
         31 . The population of cells of  claim 26 , comprising at least two different cell types.  
     
     
         32 . The population of cells of  claim 31 , wherein the cell types are neural progenitor cells and glial cells.  
     
     
         33 . The population of cells of  claim 32 , wherein the glial cells express the cell surface glycoprotein CD44.  
     
     
         34 . The population of cells of  claim 27 , wherein the cells are isolated from embryonic pig between about days 30 and 40 of gestation.  
     
     
         35 . The population of cells of  claim 26 , further comprising at least one of the agents or factors selected from the group consisting of an angiogenic factor, a neurotrophic factor, an antiapoptotic factor, an antioxidant, and an agent which inhibits nitric oxide production.  
     
     
         36 . The population of cells of  claim 35 , wherein the angiogenic factor is selected from the group consisting of vascular endothelial growth factor, platelet-derived growth factor, fibroblast growth factor, and epidermal growth factor.  
     
     
         37 . The population of cells of  claim 35 , wherein the neurotrophic factor is selected from the group consisting of glial cell line-derived growth factor, brain-derived neurotrophic factor, ciliary neurotrophic factor, midkine, insulin-like growth factor I and II, insulin, fibroblast growth factor, neurotrophin-3, neurotrophin 4/5, transforming growth factorβ, FK506, and cyclosporine A  
     
     
         38 . The population of cells of  claim 35 , wherein the antiapoptotic factor is selected from the group consisting of the bcl-2 gene product, the bcl-xL gene product, the bcl-Xβ gene product, the crmA gene product, nerve growth factor, platelet-derived growth factor, superoxide dismutase, catalase, and N-acetyl cysteine.  
     
     
         39 . The population of cells of  claim 35 , wherein the antioxidant is selected from the group consisting of superoxide dismutase, glutathione peroxidase, N-acetyl cysteine, a lazaroid, vitamin C, vitamin E, and beta carotene.  
     
     
         40 . The population of cells of  claim 35 , wherein the agent which inhibits nitric oxide production is selected from the group consisting of gangliosides, FK506, cyclosporine A, and superoxide dismutase.  
     
     
         41 . An isolated population of porcine striatal cells which, in unmodified form, have at least one antigen on the cell surface which is capable of stimulating an immune response against the cell in a xenogeneic subject, wherein the antigen on the cell surface is altered to inhibit rejection of the cell when introduced into the xenogeneic subject.  
     
     
         42 . The population of cells of  claim 41 , wherein the antigen on the cell surface which is altered is an MHC class I antigen.  
     
     
         43 . The population of cells  claim 42  , which is contacted prior to transplantation into a xenogeneic subject with at least one anti-MHC class I antibody, or fragment or derivative thereof, which binds to the MHC class I antigen on the cell surface but does not activate complement or induce lysis of the cell.  
     
     
         44 . The population of cells of  claim 41 , comprising embryonic porcine striatal cells.  
     
     
         45 . The population of cells of  claim 44 , which is obtained from an embryonic pig between about days 30 and 40 of gestation.  
     
     
         46 . The population of cells of  claim 45 , which is obtained from a lateral ganglionic eminence of porcine striatum.  
     
     
         47 . A porcine striatal cell isolated from a pig which is essentially free from organisms or substances which are capable of transmitting infection or disease to a xenogeneic recipient of the cells.  
     
     
         48 . The porcine striatal cell of  claim 47  which is isolated from a pig which is essentially free from at least one organism selected from the group consisting of parasites, bacteria, mycoplasma, and viruses.  
     
     
         49 . The porcine striatal cell of  claim 47 , which is an embryonic striatal cell.  
     
     
         50 . The porcine striatal cell of  claim 49 , which is obtained from an embryonic pig between about days 30 and 40 of gestation.  
     
     
         51 . The porcine striatal cell of  claim 50 , which is obtained from a lateral ganglionic eminence of porcine striatum.  
     
     
         52 . A cell culture comprising porcine striatal cells wherein at least about 30% to about 50% of the cells comprise striatal cells.  
     
     
         53 . The cell culture of  claim 52 , wherein at least about 50% of the cells are glial cells.  
     
     
         54 . The cell culture of  claim 52 , wherein the porcine striatal cells are embryonic porcine striatal cells obtained from a lateral ganglionic eminence of porcine striatum.  
     
     
         55 . The cell culture of  claim 52 , further comprising at least one of the agents or factors selected from the group consisting of an angiogenic factor, a neurotrophic factor, an antiapoptotic factor, an antioxidant, and an agent which inhibits nitric oxide production.  
     
     
         56 . An isolated porcine striatal cell which, upon transplantation into a xenogeneic subject, extends at least one neural process to at least one region of the brain of the subject.  
     
     
         57 . The porcine striatal cell of  claim 56 , wherein the region comprises a striatal efferent target region.  
     
     
         58 . The porcine striatal cell of  claim 57 , wherein the striatal efferent target region comprises substantia nigra or globus pallidus.  
     
     
         59 . A method for treating neurological deficits due to neurodegeneration in the brain of a xenogeneic subject comprising introducing into an area of neurodegeneration in the brain of a xenogeneic subject, porcine striatal cells, which, in unmodified form, have at least one antigen on the cell surface which is capable of stimulating an immune response against the cell in a xenogeneic subject, wherein the antigen on the cell surface is altered to inhibit rejection of the cell when introduced into the xenogeneic subject.  
     
     
         60 . The method of  claim 59 , wherein the striatal cells are embryonic striatal cells.  
     
     
         61 . The method of  claim 59 , wherein the area of neurodegeneration is in the substantia nigra.  
     
     
         62 . The method of  claim 61 , wherein the subject is a human with Huntington's disease.  
     
     
         63 . The method of  claim 59 , wherein the porcine striatal cells are contacted prior to introduction into the subject with at least one molecule which binds to at least one antigen on the cell surface which is capable of stimulating an immune response against the cell in the subject to alter the antigen on the cell surface to inhibit rejection of the cell when introduced into a subject.  
     
     
         64 . The method of  claim 63 , wherein the antigen on the surface of the porcine striatal cells which is altered is an MHC class I antigen.  
     
     
         65 . The method of  claim 64 , wherein the porcine striatal cells are contacted prior to introduction into a subject with at least one anti-MHC class I antibody, or fragment or derivative thereof, which binds to the MHC class I antigen on the cell surface but does not activate complement or induce lysis of the cell.  
     
     
         66 . The method of  claim 65 , wherein the anti-MHC class I antibody is an anti-MHC class I F(ab′) 2  fragment.  
     
     
         67 . The method of  claim 66 , wherein the anti-MHC class I F(ab′) 2  fragment is a F(ab′) 2  fragment of a monoclonal antibody PT85.  
     
     
         68 . The method of  claim 59 , further comprising administering an immunosuppressive agent to the subject.  
     
     
         69 . A method for treating neurological deficits due to neurodegeneration in the brain of a xenogeneic subject comprising introducing porcine striatal cells obtained from a pig which is essentially free from organisms or substances which are capable of transmitting infection or disease to the subject into an area of neurodegeneration in the brain of the subject.  
     
     
         70 . The method of  claim 69 , wherein the area of neurodegeneration is in the basal ganglia  
     
     
         71 . The method of  claim 69 , wherein the area of neurodegeneration is in the striatum of the subject.  
     
     
         72 . The method of  claim 69 , wherein the porcine striatal cells are embryonic striatal cells.  
     
     
         73 . The method of  claim 72  , wherein the striatal cells are obtained from an embryonic pig between about days 30 and 40 of gestation.  
     
     
         74 . The method of  claim 73 , wherein the striatal cells are obtained from a lateral ganglionic eminence of porcine striatum.  
     
     
         75 . The method of  claim 69 , wherein the subject is a human with Huntington's disease.  
     
     
         76 . A method for treating neurological deficits due to neurodegeneration in the brain of a xenogeneic subject comprising introducing porcine striatal cells into an area of neurodegeneration in the brain of a xenogeneic subject.  
     
     
         77 . The method of  claim 76 , wherein the area of neurodegeneration is in the basal ganglia of the subject.  
     
     
         78 . The method of  claim 76 , wherein the area of neurodegeneration is in the striatum of the subject.  
     
     
         79 . The method of  claim 76 , wherein the subject is a human with Huntington's disease.  
     
     
         80 . The method of  claim 76 , wherein the porcine striatal cells are embryonic striatal cells.  
     
     
         81 . The method of  claim 80 , wherein the porcine striatal cells are obtained from a lateral ganglionic eminence of porcine striatum.  
     
     
         82 . The method of  claim 76 , further comprising administering at least one of the agents or factors selected from the group consisting of an angiogenic factor, a neurotrophic factor, an antiapoptotic factor, an antioxidant, and an agent which inhibits nitric oxide production.  
     
     
         83 . The method of  claim 82 , wherein the angiogenic factor is selected from the group consisting of vascular endothelial growth factor, platelet-derived growth factor, fibroblast growth factor, and epidermal growth factor.  
     
     
         84 . The method of  claim 82  , wherein the neurotrophic factor is selected from the group consisting of glial cell line-derived growth factor, brain-derived neurotrophic factor, ciliary neurotrophic factor, midkine, insulin-like growth factor I and II, insulin, fibroblast growth factor, neurotrophin-3, neurotrophin 4/5, transforming growth factor β, FK506, and cyclosporine A  
     
     
         85 . The method of  claim 82 , wherein the antiapoptotic factor is selected from the group consisting of the bcl-2gene product, the bcl-xL gene product, the bcl-Xβ gene product, the crmA gene product, nerve growth factor, platelet-derived growth factor, superoxide dismutase, catalase, and N-acetyl cysteine.  
     
     
         86 . The method of  claim 82 , wherein the antioxidant is selected from the group consisting of superoxide dismutase, glutathione peroxidase, N-acetyl cysteine, a lazaroid, vitamin C, vitamin E, and beta carotene.  
     
     
         87 . The method of  claim 87 , wherein the agent which inhibits nitric oxide production is selected from the group consisting of gangliosides, FK506, cyclosporine A, and superoxide dismutase.  
     
     
         88 . A method for treating Huntington's disease in a human subject comprising introducing cells obtained from a lateral ganglionic eminence of embryonic porcine striatum into the brain of the human subject.  
     
     
         89 . The method of  claim 88 , wherein the cells are introduced into an area of striatal neurodegeneration in the brain of the human subject.  
     
     
         90 . A method for treating epilepsy in a human subject comprising introducing cells obtained from a lateral ganglionic eminence of embryonic striatum into the brain of the human subject.  
     
     
         91 . A method for isolating neural cells from a lateral ganglionic eminence of embryonic porcine striatum comprising dissecting the lateral ganglionic eminence from the medial ganglionic eminence of the striatum of an embryonic pig between about 30 to 40 days of gestation and isolating the neural cells from the lateral ganglionic eminence.  
     
     
         92 . The porcine striatal cell of  claim 1 , which is derived from a porcine stem cell.  
     
     
         93 . The composition of  claim 16 , wherein the porcine striatal cell is derived from a porcine stem cell.  
     
     
         94 . The porcine striatal cell of  claim 22 , which is derived from a porcine stem cell.  
     
     
         95 . The porcine striatal cell of  claim 47 , which is derived from a porcine stem cell.  
     
     
         96 . The method of  claim 76 , wherein the porcine striatal cells are derived from porcine stem cells.