Abstract:

To isolate and culture the stem  cells from human glioblastoma  SC 326 and SC 189 cells and to investigate the biological characteristics such as  self-renewal,proliferation,and differentiation.Methods The human glioblastoma SC 326 and SC 189 cells were cultured in serum-free media,the capability of forming neurospheres was detected.Clone forming rate assay was performed to assess the capacity of self-renewal and clonogenic potential of the cells.Immunofluorescence staining method was used to study the expressions of surface markers of stem cells and the multi-differentiation function of neurospheres.Results The cells isolated from SC 326 and SC 189 cultured in serum-free media for 24 h had strong characteristics of self-renewal,proliferation,and neurosphere reformation.The  proliferation rates of SC 326 were (2.10±4.33)%,(4.03±4.14)%, and (6.91±5.12)%; and those of SC 189 were (12.79±2.32)%,(15.78±3.25)%, and (37.91±4.58)% in the first three generations; the rates of neurospheres formation of GSC 326 and GSC 189 single cells were 5.56% and 8.33%,and the rates of sub-spheres formation were 9.89% and 14.58%; the maximum clone formation rates of   GSC 326 P3 and P7 neurospheres were (12.67±2.86)% and (20.44±1.73)%,and those of  GSC 189 P3 and P7 were (32.00±1.00)% and (42.67±5.03)%.Immunofluorescence staining showed that the  neuron stem cells marker CD133 and nestin   positively  expressed;and  the glial fibrillary acidic protein(GFAP) and neuron-specific class β-tubulin Ⅲ(β-Tubulin Ⅲ) weekly expressed.After  treated with differentiated media,the expressions of differentiation markers β-Tubulin Ⅲ and GFAP were  positive.Conclusion A few cells existed in glioblastoma SC 326 and SC 189 cell lines in the primary and early stage have the capacities of self-renewal,proliferation and multi-differentiation and the cells present definite features of stem cells.
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Key words: glioblastoma stem cells, self-renewal, proliferation, differentiation