earticle

영어

Cell surface gangliosides, which exist in glycosphingolipids (GSLs)-enriched domains, play important regulatory roles in cell proliferation and differentiation .Recently, gangliosides are known as important role in growth of neuronal progenitor cells. In this study, the relationship between gangliosides expression and neuronal cell development was investigated using an in vitro model of neuronal differentiation from human mesenchymal stem cells (hMSCs) that are able to differentiate into a range of specific cell types in vitro and in vivo. First, to verify the isolated population contained pure hMSCs, we performed positive and negative characterization by fluorescence- activated cell sorting (FACs) analysis, thus we identified pure hMSCs. Next, hMSCs were differentiated into neuronal cell withα-MEM + 20%FBS + 1mM β-mercaptoethanol (BME) for 24h and transferred to serum free α-MEM + 2mM BME for 5 hour. Neuronal induced cells maintenance with α-MEM + 10%FBS + 2% DMSO +200uM β- hydroxyanisole (BHA) + 10ng/mL bFGF + 10ng/mL EGF + 25ng/mL NGF for 2weeks. High-performance thin-layer chromatography (HPTLC) showed that gangliosides GM3, GD3 and GD1a expressed in differentiated into neuronal cells for 2weeks, especially GD3 increased comparison to control cells. Immunofluorescence staining also agreed with the results of HPTLC assay. These differentially expressed gangliosides suggest that gangliosides may have specific functions in stem cells and during neuronal differentiation. Therefore, these result also suggest that regulation of gangliosides expression have used as the marker for differentiated neuronal cell from hMSCs.