The possible cytotoxic effects of vancomycin and its complex with beta-cyclodextrin (β-CD) on human glial cell line (CRL 8621) were studied accordingly by means of MTS assay. The cultured cells were incubated with various concentrations of vancomycin, β-CD as well as β-CD/vancomycin complex ranging from 4.69 to 300 ug/ml. A linear dose-dependency cytotoxicity followed by hermetic-like biphasic dose-dependence was observed after incubation period of 72 hours. In general, significant increase (p<0.001) of cell proliferation was observed at lower concentrations: <18.75 μg/ml for cells treated with β-CD and their complex while < 9.38 μg/ml for cells treated with vancomycin. In contrary, regardless of the treatments given, significant (p<0.001) reduce in cell survival was found at higher concentrations >150 μg/ml. In particular, 50 % inhibitory in vitro was achieved at the concentrations of 115.95 μg/ml (for β-CD), 116.48 μg/ml (for vancomycin) and 115.44 μg/ml (for β-CD/vancomycin complex).