神经胶质瘤干细胞样细胞的分离鉴定及其生物学特性初探

Biological characteristics and isolation and identification of stem cell-like glioblastoma cells

目的分选神经胶质瘤C6细胞中的侧群(SP)细胞,探讨其相关生物学特性。方法 Hoechst33342染色流式细胞仪分选SP细胞,另取非侧群(NSP)细胞作为对照。MTT法绘制细胞生长曲线,平板克隆法检测细胞克隆形成能力,无血清悬浮培养观察细胞的球囊形成能力,裸鼠移植瘤实验检测细胞的成瘤能力,流式细胞仪检测细胞CD133的表达。结果神经胶质瘤C6细胞中SP细胞的比例为(4.24±1.36)%。SP细胞的生长速度显著快于NSP细胞(P<0.05)。SP细胞在无血清条件下培养有形成球囊的特性,而NSP细胞无球囊性生长的特性。SP细胞的克隆形成率为(31.24±6.48)%显著高于NSP细胞的(9.63±1.16%)(P<0.01)。103、104、105、106个SP细胞接种裸鼠皮下,成瘤率分别为40.00%、60.00%、80.00%和100.00%,而NSP细胞的成瘤率分别为0、0、60.00%和70.00%,两组之间差异有显著性(P<0.05)。SP细胞CD133表达的量为(61.74±7.32)%显著高于NSP的(1.06±0.81)%(P<0.01)。结论神经胶质瘤中存在SP细胞,SP细胞高表达干细胞标志蛋白CD133,SP细胞分选可以富集神经胶质瘤肿瘤干细胞样细胞。

【Objective】 To study the biological characteristics of side population（SP） cells sorted from glioblastoma C6 cells.【Methods】 Hoechst33342 staining and flow cytometric sorter were used to sort SP and NSP cells.MTT assay was used to draw the growth curve.The colony-formation ability was detected by plate colony assay.The serum-free medium was used to form cell sphere.The xenograft model was established by nude mices to analyze the oncogenicity of SP and NSP cells.The expression of CD133 was determined by flow cytometry analysis.【Results】 The SP cells in glioblastoma C6 cells were（14.24±1.36）%.The proliferation ability,colony-formation ability and oncogenicity of SP cells were higher than that of NSP cells（P 〈0.05）.Compared with NSP cells,SP cells could form cell sphere.The expression of CD133 in SP cells was（61.74±7.32）%,and which was higher than that in NSP cells（P 〈0.01）.【Conclusion】 The SP cells sorted from glioblastoma C6 cells could enrich tumor stem cells.