The glutamate clearance function of adipose stromal cells-derived astrocytes

Abstract

ADSCs-derived astrocytes qualify the morphology, ultrastructure and membrane electrical potential, which are all unique to astrocytes. But whether they have the glutamate clearance function like mature astrocytes is under exploration. ADSCs were extracted, cultured and induced into astrocytes for 48 h, 7d, 14d and 21d in vitro. Inverted phase contrast microscope was used to observe the morphology of the cells in each group. Immunocytochemistry assay, immunofluorescence assay and Western blotting were used to detect the expression of GFAP, EAAT2 and GS of the cells in each group. The cells were cultured in glutamate solution for 1, 2, 3 and 4 h respectively before the solution collected. The glutamate concentration of the solution was detected using Glutamate Colorimetric Assay Hit. ADSCs-derived astrocytes expressed GFAP, EAAT2 and GS, all of which increased gradually and reached peak when induced for 14 days. In induction for 48 h, 7d and 14d groups, the extracellular glutamate concentration decreased gradually during the cells cultured in glutamate solution for 1, 2, 3 and 4 h, among which the decrease extent was most prominent in 14d group, while the extracellular glutamate concentration had no change in uninduction and induction for 21d group. ADSCs-derived astrocytes expressed EAAT2 and GS, meanwhile had the function of clearing glutamate, which was prominent when induced into astrocytes for 7–14 days.