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 embryonicderived 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 BDNFtreated 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.

mfat‐1 transgene protects cultured adult neural stem cells against cobalt chloride‐mediated hypoxic injury by activating Nrf2/ARE pathways

“…Because of their many advantages (Giusto et al, ), adult NSCs have been regarded as an excellent source of replacement therapy for human ischemic stroke. However, the primary obstacle to clinical application is the loss of physiological function of endogenous NSCs and exogenous adult NSCs resulting from hypoxic‐ischemic injury in the ischemic penumbra (Azevedo‐Pereira & Daadi, ; Bazan et al, ; Rosenblum et al, ). Our results provide valuable information for the development of effective adult NSC replacement therapy for ischemic stroke.…”

“…In the ideal scenario, soon after ischemic stroke, endogenous or exogenous adult NSCs would exhibit proliferation, migration, and differentiation to repair neural function damage (Chung et al, ; Cramer, ). However, because of the loss of nutrients and oxygen in the ischemic penumbra, the majority of newly generated NSCs die soon after stroke, and their physiological function is lost (Azevedo‐Pereira & Daadi, ; Bazan, Marcheselli, & Cole‐Edwards, ; Rosenblum et al, ). Therefore, figuring out how to protect adult NSCs against hypoxic‐ischemic injury after ischemic stroke is key to effective adult NSC replacement therapy.…”

Opportunities and Challenges: Stem Cell‐Based Therapy for the Treatment of Ischemic Stroke

“…Heat shock was reported to increase Sca1+ stem cells survival in ischemic heart [61]. BDNF pretreatment enhanced NSCs survival at the brain cortex in hypoxia-ischemia mice [62]. In addition, gene modification was another promising strategy to enhance transplanted stem cells survival.…”

Insight on stem cell preconditioning and instructive biomaterials to enhance cell adhesion, retention, and engraftment for tissue repair

“…One of the effective ways to enhance the success of cell therapy is to engineer the cells with plasmids encoding for signaling molecules such as cell survival molecules, growth factors, and cytokines [149,150]. Such manipulations have increased the survival of transplanted cells, and modulated their paracrine behavior for enhanced expression of bioactive growth factors or cytokines [151].…”