title: Bone marrow stromal cells rescue ischemic brain by trophic effects and phenotypic change toward neural cells.
abstract: Background. Transplantation of bone marrow stromal cells (BMSCs) may contribute to functional recovery after stroke. This study was designed to clarify their mechanisms, trophic effects of neurotrophic factors, and neural differentiation. Methods. Mouse neurons exposed to glutamate were cocultured with mouse BMSCs. Either neutralizing antibodies against brain-derived neurotrophic factor (BDNF) or nerve growth factor (NGF) or Trk inhibitor K252a was added to explore the mechanism of their protective effects. Fluorescence in situ hybridization (FISH) was used to assess BDNF or NGF mRNA expression in BMSCs. The mice were subjected to permanent focal ischemia, and 7 days later, either BMSCs or the vehicle was stereotactically transplanted into the ipsilateral striatum. The mouse brains were processed for FISH and immunostaining 2 or 4 weeks after transplantation. Results. BMSCs significantly ameliorated glutamate-induced neuronal death. Treatment with anti-BDNF antibody significantly reduced their protective effects. FISH analysis showed that the majority of BMSCs expressed BDNF and NGF mRNA in vitro. BMSC transplantation significantly improved the survival of neurons in peri-infarct areas. FISH analysis revealed that approximately half of BMSCs expressed BDNF and NGF mRNA 2 weeks after transplantation; however, the percentage of BDNF and NGF mRNA-positive cells decreased thereafter. Instead, the percentage of microtubule-associated protein 2-positive BMSCs gradually increased during 4 weeks after transplantation. Conclusions. These findings strongly suggest that BDNF may be a key factor underlying the trophic effects of BMSCs. BMSCs might exhibit the trophic effect in the early stage of cell therapy and the phenotypic change toward neural cells thereafter.
authors.affiliation: Depertment of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Shichinohe
authors.affiliation: Shionogi Innovation Center for Drug, Shionogi & Co Ltd, Sapporo, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Ishihara
authors.affiliation: Shionogi Innovation Center for Drug, Shionogi & Co Ltd, Sapporo, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Takahashi
authors.affiliation: Shionogi Innovation Center for Drug, Shionogi & Co Ltd, Sapporo, Japan.
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Tanaka
authors.affiliation: Depertment of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Michiyuki
authors.initials: M
authors.lastname: Miyamoto
authors.affiliation: Depertment of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Yamauchi
authors.affiliation: Depertment of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Hisayasu
authors.initials: H
authors.lastname: Saito
authors.affiliation: Shionogi Innovation Center for Drug, Shionogi & Co Ltd, Sapporo, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Takemoto
authors.affiliation: Depertment of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Kiyohiro
authors.initials: K
authors.lastname: Houkin
authors.affiliation: Department of Neurosurgery, Graduate School of Medicine and Pharmaceutical Science for Education, University of Toyama, Toyama, Japan skuroda@med.u-toyama.ac.jp.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kuroda
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism*; D002452:Cell Count; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D018920:Coculture Techniques; D004195:Disease Models, Animal; D004622:Embryo, Mammalian; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D020244:Infarction, Middle Cerebral Artery / Q000601:surgery*; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D020932:Nerve Growth Factor / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D008869:Microtubule-Associated Proteins; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D020932:Nerve Growth Factor; D010751:Phosphopyruvate Hydratase
keywords: bone marrow stromal cells; coculture; neurotrophic factor; permanent middle cerebral artery occlusion; regenerative medicine