前列腺癌干细胞的分离及鉴定

Isolation and Preliminary Characterization of Prostate Cancer Stem Cells

前列腺癌DU145细胞在普通培养基中培养至对数生长期,无血清培养基重悬细胞培养至形成球状体,以细胞三维(3D)培养和二维(2D)培养方法分离前列腺癌类干细胞,观察细胞球的形成及其生长情况,通过CCK-8细胞增殖实验和细胞划痕修复实验检测所分离细胞的增殖能力,结果显示前列腺癌DU145细胞在普通培养基中呈长梭形贴壁生长,无血清培养基培养72 h形成大量细胞球聚集体,细胞3D培养分离的细胞球的透明度和形态比2D培养分离的细胞好,3D培养分离细胞的增殖能力和划痕修复能力明显强于2D培养分离细胞。因此,细胞3D培养技术联合无血清培养基有利于前列腺癌干细胞的分离。

Prostate cancer DU145 cells were cultured in a normal medium to logarithmic growth phase.Cells were resuspended in serum-free medium until spheroid was formed.Three-dimensional(3D)culture and two-dimensional(2D)culture methodswere used to separate prostate cancer stem cells,respectively.The shapes and growth rates of the cell spheres were observed,meanwhile the proliferation capabilities of the separated cells were tested using CCK-8 assay kits and cell Wound-healing experiments.The results showed that prostate cancer DU145 cells adhered on the substrate with spindle shape in normal medium,leading to a large number of cell spheres in serum-free medium after 72 hours culture.The transparency and morphology of cell spheres isolated from 3D culture were better than those of cells isolated from 2D culture.In addition,the proliferation abilities and Wound-healing abilities of cells isolated from 3D culture were significantly stronger than those of cells isolated from 2D culture.Therefore,3D culture technology cooperated with serum-free medium was benefit for separation of prostate cancer stem cells in vitro.