title: BDNF Pretreatment of Human Embryonic-Derived Neural Stem Cells Improves Cell Survival and Functional Recovery After Transplantation in Hypoxic-Ischemic Stroke.
abstract: Intra-arterial neural stem cell (NSC) therapy has the potential to improve long-term outcomes after stroke. Here we evaluate if pretreatment of NSCs with brain-derived neurotrophic factor (BDNF) prior to transplantation improves cell engraftment and functional recovery following hypoxic-ischemic (HI) stroke. Human embryonic-derived NSCs with or without BDNF pretreatment (1 h, 100 ng/ml) were transplanted 3 days after HI stroke. Functional recovery was assessed using the horizontal ladder test. Cell engraftment was evaluated using bioluminescence imaging (BLI) and histological counts of SC121(+) cells. Fluoro-Jade C (FJC) and NeuN stains were used to evaluate neuroprotection. The effect of BDNF on NSCs was analyzed using a migration assay, immunocytochemistry, Luminex proteomic assay, and RT-qPCR.BLI analysis demonstrated significantly higher photon flux in the BDNF-treated NSC group compared to untreated NSC (p = 0.049) and control groups (p = 0.0021) at 1 week after transplantation. Immunohistochemistry confirmed increased transplanted cell survival in the cortex (p = 0.0126) and hippocampus (p = 0.0098) of animals injected with BDNF-treated NSCs compared to untreated NSCs. Behavioral testing revealed that the BDNF-treated NSC group demonstrated increased sensorimotor recovery compared to the untreated NSC and control groups (p < 0.001) over the 1-month period (p < 0.001) following transplantation. A significant improvement in performance was found in the BDNF-treated NSC group compared to the control group at 14, 21, and 28 (p < 0.05) days after transplantation. The cortex and hippocampus of the BDNF-treated NSC group had significantly more SC121(+) NSCs (p = 0.0125, p = 0.0098), fewer FJC(+) neurons (p = 0.0370, p = 0.0285), and a higher percentage of NeuN(+) expression (p = 0.0354) in the cortex compared to the untreated NSC group. BDNF treatment of NSCs resulted in significantly greater migration to SDF-1, secretion of M-CSF, VEGF, and expression of CXCR4, VCAM-1, Thrombospondins 1 and 2, and BDNF. BDNF pretreatment of NSCs results in higher initial NSC engraftment and survival, increased neuroprotection, and greater functional recovery when compared to untreated NSCs.
authors.affiliation: Department of Neurosurgery, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Sahar
authors.initials: S
authors.lastname: Rosenblum
authors.forename: Tenille N
authors.initials: TN
authors.lastname: Smith
authors.forename: Nancy
authors.initials: N
authors.lastname: Wang
authors.forename: Joshua Y
authors.initials: JY
authors.lastname: Chua
authors.forename: Erick
authors.initials: E
authors.lastname: Westbroek
authors.forename: Kendrick
authors.initials: K
authors.lastname: Wang
authors.forename: Raphael
authors.initials: R
authors.lastname: Guzman
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology*; D015815:Cell Adhesion Molecules / Q000096:biosynthesis; D002454:Cell Differentiation; D002460:Cell Line; D002465:Cell Movement; D002470:Cell Survival; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D002540:Cerebral Cortex / Q000166:cytology; D019743:Chemokines, CXC / Q000096:biosynthesis; D053595:Embryonic Stem Cells / Q000166:cytology / Q000637:transplantation*; D057809:HEK293 Cells; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D020925:Hypoxia-Ischemia, Brain / Q000473:pathology / Q000628:therapy*; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D058953:Neural Stem Cells / Q000166:cytology / Q000637:transplantation*; D019707:Receptors, Chemokine / Q000096:biosynthesis; D020127:Recovery of Function; D020521:Stroke / Q000473:pathology / Q000628:therapy*
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D015815:Cell Adhesion Molecules; D019743:Chemokines, CXC; D019707:Receptors, Chemokine; C579455:BDNF protein, human