Platelet-derived growth factor-responsive neural precursor cells and progeny thereof

Abstract

This invention provides platelet-derived growth factor-responsive neural precursor (PRP) cells and methods of producing such cells in vivo or in vitro. These cells can further be used to generate neurons, oligodendrocytes and/or astrocytes.

1 .- 52 . (canceled) 
   
   
       53 . An isolated clonally expanded or self-renewed population of mammalian PDGF-responsive neural precursor (PRP) cells that express PDGF receptor alpha produced by a method comprising culturing brain tissue in a culture medium containing PDGF under conditions allowing clonal proliferation or differentiation of the PRP cells. 
   
   
       54 . A method of increasing PRP cell numbers in a mammal, comprising administering a PDGFR agonist to the mammal in an effective amount for intracranial delivery of the PDGFR agonist to increase PRP cell numbers. 
   
   
       55 . The method of  claim 54 , wherein the PDGFR agonist compromises PDGF. 
   
   
       56 . The method of  claim 54 , wherein the mammal does not receive EGF or FGF. 
   
   
       57 . The method of  claim 54 , further compromising administering FGF2, BDNF or NT-3 substantially simultaneously with the PDGFR agonist to the mammal. 
   
   
       58 . The method of  claim 54 , wherin the PDGFR agonist is administered to the brain of the mammal. 
   
   
       59 . The method of any of  claim 54 , wherein the PDGFR agonist is administered locally, regionally, or systemically. 
   
   
       60 . The method of  claim 54 , wherein the PDGFR agonist is administered intracranially, intravenously, intravascularly, intramuscularly, subcutaneously, intraperitoneally, topically, orally, nasally or by inhalation. 
   
   
       61 . (canceled) 
   
   
       62 . A method of producing oligodendrocytes, comprising:
 (a) culturing brain tissue from a mammal in a culture medium comprising a PDGFR agonist and allowing proliferation of PRP cells; and   (b) differentiating the proliferated PRP cells to produce oligodendrocytes.   
   
   
       63 . The method of  claim 62 , wherein step (b) is performed by contacting the proliferated PRP cells with an effective amount of thyroid hormone or T3. 
   
   
       64 . The method of  claim 62 , further compromising contacting the oligodendrocytes with an effective amount of BMP-2 and CNTF to produce neurons and astrocytes. 
   
   
       65 . The method of  claim 62 , further compromising clonally expanding the proliferated PRP cells by contacting said cells with PDGF and FGF-2; or PDGF and BDNF; or PDGF and NT-3 prior to step (b). 
   
   
       66 . A method of producing neurons, comprising: (a) culturing brain tissue from a mammal in a culture medium comprising PDGFR agonist and allowing proliferation of PRP cells; and (b) differentiating the proliferated PRP cells to produce neurons. 
   
   
       67 . The method of  claim 66 , wherein step (b) is performed by contacting the proliferated PRP cells with an effective amount of BMP-2. 
   
   
       68 . The method of  claim 66 , further comprising clonally expanding the proliferated PRP cells by contacting said cells with PDGF and FGF-2; or PDGF and BDNF; or PDGF and NT-3 prior to step (b). 
   
   
       69 . A method of producing astrocytes, comprising:
 (a) culturing brain tissue from a mammal in a culture medium comprising PDGFR agonist and allowing proliferation of PRP cells; and   (b) differentiating the proliferated PRP cells to produce astrocytes.   
   
   
       70 . The method of  claim 69 , wherein step (b) is performed by contacting the proliferated PRP cells with an effective amount of BMP-2 and CNTF. 
   
   
       71 . The method of  claim 69 , further comprising expanding the proliferated PRP cells by contacting said cells with PDGF and FGF-2; or PDGF and BDNF; or PDGF and NT-3 prior to step (b). 
   
   
       72 .- 76 . (canceled) 
   
   
       77 . A method of increasing oligodendrocytes, neurons or astrocytes in a mammal, comprising:
 (a) administering an effective amount of PDGFR agonist to the mammal to proliferate PRP cells; and   (b) administering an effective amount of thyroid hormone or T3 to increase oligodendrocytes, BMP-2 to increase neurons, or both BMP-2 and CNTF to increase astrocytes.   
   
   
       78 . The method of  claim 77 , further comprising administering FGF2, BDNF or NT-3 substantially simultaneously with the PDGFR agonist to the mammal. 
   
   
       79 . The method of  claim 77 , wherein the mammal is not administered EGF or FGF. 
   
   
       80 . The method of  claim 77 , wherein the PDGFR agonist, thyroid hormone, T3, BMP-2 or CNTF is delivered to the cranium of the mammal. 
   
   
       81 . The method of  claim 77 , wherein the PDGFR agonist, thyroid hormone, T3, BMP-2 or CNTF is administered to the brain of the mammal. 
   
   
       82 . The method of  claim 77 , wherein the PDGFR agonist is administered locally, regionally or systemically. 
   
   
       83 . The method of  claim 77 , wherein the PDGFR agonist is administered intracranially, intravenously, intravascularly, intramuscularly, subcutaneously, intraperitoneally, topically, orally, nasally or by inhalation. 
   
   
       84 . (canceled) 
   
   
       85 . The method of  claim 77 , wherein step (a) is performed prior to step (b). 
   
   
       86 . The method of  claim 77 , wherein step (a) is performed at least one day prior, three days prior or a week prior to step (b). 
   
   
       87 . The method of  claim 77 , wherein step (a) is performed concurrently with step (b). 
   
   
       88 . The method of  claim 77 , wherein the mammal is in need of increased numbers of oligodendrocytes, neurons or astrocytes. 
   
   
       89 . The method of  claim 77 , wherein the mammal suffers from a loss of or injury to oligodendrocytes, neurons or astrocytes. 
   
   
       90 . The method of  claim 77 , wherein the mammal is afflicted with or is at risk of affliction with a neurological disease or disorder, or undesirable medical condition. 
   
   
       91 . The method of  claim 90 , wherein the neurological disease comprises a neurodegenerative disease. 
   
   
       92 . The method of  claim 90 , wherein the neurological disease or undesirable medical condition comprises a stroke, aneurysm, brain or spinal cord injury or cranium or spinal column trauma. 
   
   
       93 . The method of  claim 92 , wherein the brain or spinal cord injury, or cranium or spinal column trauma, is caused by a stroke or surgery. 
   
   
       94 . The method of  claim 93 , wherein the stroke is hemorrhagic stroke, focal ischemic stroke or global ischemic stroke. 
   
   
       95 . The method of  claim 90 , wherein the neurological disease or undesirable medical condition affects central or peripheral nerves. 
   
   
       96 . The method of  claim 95 , wherein the central nerves comprise brain or spinal cord. 
   
   
       97 . The method of  claim 95 , wherein the peripheral nerves comprise one or more of motor, sensory or autonomic nerves. 
   
   
       98 . (canceled) 
   
   
       99 . The method of treating or ameliorating a disease, disorder or undesirable medical condition associated with neuron, oligodendrocytes or astrocyte loss, injury or dysfunction, comprising administering an effective amount of PDGFR agonist to a mammal harboring the disease, disorder or medical condition, as well as one or more of FGF-2, thyroid hormone, T3, BMP-2 or CNTF. 
   
   
       100 . The method of  claim 99 , further comprising administering to the mammal one or more agents selected from PDGF; PDGF and FGF-2; PDGF and BDNF; PDGF and NT-3; thyroid hormone; T3; BMP-2; BMP-2 and CNTF. 
   
   
       101 . The method of  claim 99 , wherein the undesirable medical condition comprises a neurological injury or trauma. 
   
   
       102 . The method of  claim 101 , wherein the neurological injury or trauma affects central or peripheral nerves. 
   
   
       103 . The method of  claim 102 , wherein the central nerves comprise brain or spinal cord. 
   
   
       104 . The method of  claim 102 , wherein the peripheral nerves comprise one or more of motor, sensory or autonomic nerves. 
   
   
       105 . The method of  claim 101 , wherein the neurological injury or trauma comprises stroke, aneurysm, brain or spinal cord injury or cranium or spinal column trauma or injury. 
   
   
       106 . The method of  claim 105 , wherein the stroke is hemorrhagic stroke, focal ischemic stroke or global ischemic stroke. 
   
   
       107 . The method of  claim 99 , wherein the disease, disorder or undesirable medical condition comprises Alzheimer's Disease, multiple sclerosis (MS), macular degeneration, glaucoma, diabetic retinopathy, peripheral neuropathy, Huntington's Disease, amyotrophic lateral sclerosis (ALS), Parkinson's Disease, stroke, depression, epilepsy, neurosis or psychosis. 
   
   
       108 . (canceled) 
   
   
       109 . A method of identifying an agent that modulates clonal proliferation or self renewal or differentiation of a neural precursor cell comprising:
 (a) providing an isolated or purified mammalian platelet derived growth factor (PDGF)-responsive neural precursor (PRP) cell, wherein said cell expresses PDGF receptor alpha, and wherein said cell, when contacted with one or more of thyroid hormone, bone morphogenetic protein-2 (BMP-2), ciliary neurotrophic factor (CNTF) or triiodothyronine (T3), gives rise to a differentiated neural cell that expresses detectable amounts of one or more protein markers selected from: GABA, parvalbumin, beta-II tubulin, calbindin D, calretinin, O4, neurofilament M (NFM), myelin basic protein (MBP), TOA-64/TUC-2 and GFAP or progeny cells thereof;   (b) contacting the cell or cells of step (a) with a candidate agent; and   (c) determining if the candidate agent modulates clonal expansion or differentiation of the cell or cells.   
   
   
       110 - 111 . (canceled) 
   
   
       112 . A method of identifying an agent that modulates clonal proliferation or self renewal or differentiation of a neural precursor cell comprising:
 (a) providing an isolated or purified mammalian platelet derived growth factor (PDGF)-responsive neural precursor (PRP) cell, wherein said cell expresses PDGF receptor alpha, and wherein said cell, when contacted with one or more of thyroid hormone, BMP-2, CNTF or T3, gives rise to a differentiated neuron, oligodendrocyte, astrocyte or mixture thereof, or progeny cells thereof;   (b) contacting the cell or cells of step (a) with a candidate agent; and   (c) determining if the candidate agent modulates clonal expansion or differentiation of the cell or cells.