Evaluation of Bone Marrow- and Brain-Derived Neural Stem Cells in Therapy of Central Nervous System Autoimmunity

Abstract

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease resulting from an autoimmune response against central nervous system (CNS) myelin. Inflammatory infiltration and demyelination of the CNS are hallmarks of MS and its animal model, experimental autoimmune encephalomyelitis (EAE).1-3 MS begins when peripherally activated myelin-reactive T cells infiltrate into the CNS, followed closely by other immune cells, including naïve myelin-reactive T cells, polyclonal T cells, macrophages, dendritic cells, B cells, and neutrophils.

The Coumarin Derivative Osthole Stimulates Adult Neural Stem Cells, Promotes Neurogenesis in the Hippocampus, and Ameliorates Cognitive Impairment in APP/PS1 Transgenic Mice

“…After 7-14 d of culture, NSCs changed morphology and developed markers of neurons, astrocytes and oligodendroglia as verified by immunocytochemistry staining. 7,10) Cell Viability and Proliferation Assays Single NSCs at 5th passage were seeded at 5.0×10 4 cells/mL in proliferation media and exposed to various concentrations of Ost (0, 10, 50, and 100 µM) (34)(35)(36). After 3 d of co-culture, cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Sigma) assay as previously described.…”

“…Cell counter of ImageJ software (National Institutes of Health, Bethesda, MD, U.S.A.) was used to count the cells. 7,10) Statistical analyses were performed using SPSS 13.0 software (SPSS, Chicago, IL, U.S.A.). All data are expressed as mean±standard deviation (S.D.)…”

“…Seven micrometers frozen sections were prepared using a cryostat microtome (Leica, Nussloch, Germany). 7,10) Cryosections of the brain and NSCs maintained in growth medium or differentiation medium in vitro were fixed with 4% paraformaldyhyde plus 0.5% glutaraldehyde for 15 min, then washed twice with PBS. Sections were incubated with 10% goat serum in PBS for 30 min, after which primary antibodies were added and incubated at 4°C overnight.…”

“…ImageJ software was used for quantitative analysis of fluorescence images. 7,10) Reverse Transcription-Polymerase Chain Reaction (RT-PCR) NSCs treated with Ost in proliferation medium for 3 d were collected, and total RNA was extracted with TriZol and converted to cDNA by reverse transcriptase using a RevertAid First Strand cDNA Synthesis Kit (Thermo, Vilnius, Lithuania).…”

“…7,8) The adult CNS, especially the injured CNS, may provide a relatively non-permissive environment for engrafted NSCs. 7,[9][10][11] Even under the best circumstances, the cell survival in injured CNS has been estimated to be near 10%, 12,13) and few cells differentiate into mature neuronal phenotypes. 14,15) One of the most important reasons why those cells could not survive in the injured CNS is probably due to the sustained inflammation and oxidation in the disease areas.…”