title: Pre-differentiation of human neural stem cells into GABAergic neurons prior to transplant results in greater repopulation of the damaged brain and accelerates functional recovery after transient ischemic stroke.
abstract: BACKGROUND
  Despite attempts to prevent brain injury during the hyperacute phase of stroke, most sufferers end up with significant neuronal loss and functional deficits. The use of cell-based therapies to recover the injured brain offers new hope. In the current study, we employed human neural stem cells (hNSCs) isolated from subventricular zone (SVZ), and directed their differentiation into GABAergic neurons followed by transplantation to ischemic brain.
  
  
  METHODS
  Pre-differentiated GABAergic neurons, undifferentiated SVZ-hNSCs or media alone were stereotaxically transplanted into the rat brain (n=7/group) 7 days after endothelin-1 induced stroke. Neurological outcome was assessed by neurological deficit scores and the cylinder test. Transplanted cell survival, cellular phenotype and maturation were assessed using immunohistochemistry and confocal microscopy.
  
  
  RESULTS
  Behavioral assessments revealed accelerated improvements in motor function 7 days post-transplant in rats treated with pre-differentiated GABAergic cells in comparison to media alone and undifferentiated hNSC treated groups. Histopathology 28 days-post transplant indicated that pre-differentiated cells maintained their GABAergic neuronal phenotype, showed evidence of synaptogenesis and up-regulated expression of both GABA and calcium signaling proteins associated with neurotransmission. Rats treated with pre-differentiated cells also showed increased neurogenic activity within the SVZ at 28 days, suggesting an additional trophic role of these GABAergic cells. In contrast, undifferentiated SVZ-hNSCs predominantly differentiated into GFAP-positive astrocytes and appeared to be incorporated into the glial scar.
  
  
  CONCLUSIONS
  Our study is the first to show enhanced exogenous repopulation of a neuronal phenotype after stroke using techniques aimed at GABAergic cell induction prior to delivery that resulted in accelerated and improved functional recovery.
authors.affiliation: Neurotrauma Research Team, Department of Medicine, University of Melbourne, Level 4, Clinical Sciences Building, 29 Regent Street, Fitzroy, VIC, 3065, Australia. himaa@student.unimelb.edu.au.|Department of Surgery, University of Melbourne, Melbourne, VIC, Australia. himaa@student.unimelb.edu.au.
authors.forename: Hima C S
authors.initials: HC
authors.lastname: Abeysinghe
authors.affiliation: Neurotrauma Research Team, Department of Medicine, University of Melbourne, Level 4, Clinical Sciences Building, 29 Regent Street, Fitzroy, VIC, 3065, Australia. laita.bokhari@unimelb.edu.au.|Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Melbourne, VIC, Australia. laita.bokhari@unimelb.edu.au.
authors.forename: Laita
authors.initials: L
authors.lastname: Bokhari
authors.affiliation: ARC Centre of Excellence for Electromaterials Science, Intelligent Polymer Research Institute, AIIM Facility, Innovation Campus, University of Wollongong, Squires Way, Fairy Meadow, NSW 2519, Australia. anita.quigley@svhm.org.au.
authors.forename: Anita
authors.initials: A
authors.lastname: Quigley
authors.affiliation: Department of Obstetrics and Gynecology, National University of Singapore, Singapore, Singapore. mahesh_a_choolani@nuhs.edu.sg.
authors.forename: Mahesh
authors.initials: M
authors.lastname: Choolani
authors.affiliation: Department of Obstetrics and Gynecology, National University of Singapore, Singapore, Singapore. jerrychan@nus.edu.sg.
authors.forename: Jerry
authors.initials: J
authors.lastname: Chan
authors.affiliation: Cytoprotection Pharmacology Program, Centre for Eye Research, The Royal Eye and Ear Hospital Melbourne, Melbourne, VIC, Australia. g.dusting@unimelb.edu.au.|Department of Opthamology, Faculty of Medicine, University of Melbourne, Melbourne, VIC, Australia. g.dusting@unimelb.edu.au.
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Dusting
authors.affiliation: Department of Surgery, University of Melbourne, Melbourne, VIC, Australia. jcrook@uow.edu.au.|ARC Centre of Excellence for Electromaterials Science, Intelligent Polymer Research Institute, AIIM Facility, Innovation Campus, University of Wollongong, Squires Way, Fairy Meadow, NSW 2519, Australia. jcrook@uow.edu.au.|Illawarra Health and Medical Research Institute, University of Wollongong, Wollongong, NSW 2522, Australia. jcrook@uow.edu.au.
authors.forename: Jeremy M
authors.initials: JM
authors.lastname: Crook
authors.affiliation: ARC Centre of Excellence for Electromaterials Science, Intelligent Polymer Research Institute, AIIM Facility, Innovation Campus, University of Wollongong, Squires Way, Fairy Meadow, NSW 2519, Australia.
authors.forename: Nao R
authors.initials: NR
authors.lastname: Kobayashi
authors.affiliation: Neurotrauma Research Team, Department of Medicine, University of Melbourne, Level 4, Clinical Sciences Building, 29 Regent Street, Fitzroy, VIC, 3065, Australia. carlir@unimelb.edu.au.|Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Melbourne, VIC, Australia. carlir@unimelb.edu.au.
authors.forename: Carli L
authors.initials: CL
authors.lastname: Roulston
mesh_terms: D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000473:pathology; D059330:GABAergic Neurons / Q000378:metabolism / Q000637:transplantation*; D020244:Infarction, Middle Cerebral Artery / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D002546:Ischemic Attack, Transient / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D008297:Male; D009043:Motor Activity; D058953:Neural Stem Cells / Q000502:physiology*; D055495:Neurogenesis; D017208:Rats, Wistar; D020127:Recovery of Function