Abstract
Enhancement of endogenous neurogenesis after ischemic stroke may improve functional recovery. We previously demonstrated that medium B, which is a combination with epidermal growth factor (EGF) and fibronectin, can promote neural stem/progenitor cell (NSPC) proliferation and migration. Here, we showed that medium B promoted proliferation and migration of cultured NSPCs onto various 3-dimentional structures. When rat cortical neurons with oxygen glucose deprivation (OGD) were co-cultured with NSPCs, medium B treatment increased neuronal viability and reduced cell apoptosis. In a rat model with transient middle cerebral artery occlusion (MCAO), post-insult intraventricular medium B treatment enhanced proliferation, migration, and neuronal differentiation of NSPCs and diminished cell apoptosis in the infarct brain. In cultured post-OGD neuronal cells and the infarct brain from MCAO rats, medium B treatment increased protein levels of Bcl-xL, Bcl-2, phospho-Akt, phospho-GSK-3β, and β-catenin and decreased the cleaved caspase-3 level, which may be associated with the effects of anti-apoptosis. Notably, intraventricular medium B treatment increased neuronal density, improved motor function and reduced infarct size in MCAO rats. In summary, medium B treatment results in less neuronal death and better functional outcome in both cellular and rodent models of ischemic stroke, probably via promotion of neurogenesis and reduction of apoptosis.
Highlights
Www.nature.com/scientificreports and promoted the neuronal differentiation of neural stem/progenitor cell (NSPC) and diminished neuronal apoptosis in the presence of NSPCs
We previously demonstrated that medium B can enhance the proliferation and migratory capacity of NSPCs in culture[13]
With medium B treatment, NSPCs cultured on the top of Transwell membranes migrated through the small holes to the other side of the membrane, while NSPCs without medium B treatment showed a near absence of migration (Fig. 1a)
Summary
The animal experimental procedures and protocols were approved by the National Taiwan University Institutional Laboratory Animal Care Committee and the Utilization Committee (No 20140276) and AAALAC-accredited facility. All procedures met the requirements of the Animal Welfare Protection Act of the Department of Agriculture, Executive Yuan, Taiwan, developed in accord with the National Institutes of Health guide for the care and use of Laboratory animals (NIH Publications No 8023, revised 1978), and the ARRIVE (Animal Research: Reporting In Vivo Experiments) guideline. Embryos were removed from the rat, and the embryonic cerebral cortices were dissected out, washed, triturated, and cultured in the complete media containing Dulbecco’s Modified Eagle Media (DMEM)/F-12 (Gibco, Pascagoula, MS) with 1% N2 supplement (Gibco), 20 ng/ml basic fibroblast growth factor (bFGF; Invitrogen, Carlsbad, CA), and 1% antibiotic solution (Gibco) without serum. Cultures were incubated at 37 °C in a humidified atmosphere and 5% CO2 for 6 days, by which time primary neurospheres would form
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