Abstract
Objective To establish the coculture model of H2O2-injuryed astrocytes and bone marrow stromal cells (BMSCs), observe the effect of BMSCs on self-repair and survival ability of H2O2-injured astrocytes and investigate the mechanism by which BMSCs repair the brain injury.Methods H2O2-injured passage 2 astrocytes were cocultured with passage 3 BMSCs in serum-free medium as coculture group. These two kinds of cells were cultured respectively with the same cell density in same serum-free medium as astrocyte alone group and BMSCs alone group. After 3 d and 7 d cultivation, the morphological changes were observed under light microscope. After 1 d and 3 d cultivation, the supematants from the cell cultures were harvested and enzyme-linked immunosorbent assay (ELISA) method was used to quantify the levels of nerve growth factor(NGF), brain-derived neurotrophic factor(BDNF),basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF)in culture supernatants. Results After 3 d and 7 d cultivation, the astrocytes and BMSCs in coculture group grew better compared with astrocyte alone group and BMSCs alone group. The NGF,BDNF and bFGF concentrations in coculture medium were higher than the total of the concentrations in astrocyte alone medium and BMSCs alone medium after 1 d and 3 d cultivation, but the EGF concentration in coculture medium was lower than the total of the concentrations in astrocytes alone medium and BMSCs alone medium after 1 d and 3 d cultivation. Conclusions Coculture of H2O2-injured astrocytes with BMSCs can promote the self-repair ability of astrocytes and increase astrocyte survival rate.Coculture of H2O2-injured astrocytes with BMSCs can also raise NGF, BDNF and bFGF production.The therapeutic efrect of BMSCs for injured brain afcer transplantation might be in part due to the BMSCs conferred protection to astrocytes and increased neurotrophic factor production. Key words: Bone marrow stromal cells; Astrocytes; Oxidative damage; Coculture techniques; Nerve growth factors
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