星形胶质细胞在凝胶环境下的三维培养

Three-dimensional culture of astrocytes in agarose gel matrix

目的:制备不同浓度的三维琼脂糖凝胶以进行星形胶质细胞的体外培养,从而寻找适合细胞生长的理想环境。方法:分别制备1%、2%、3%的琼脂糖凝胶,利用纳米压痕仪测量其弹性模量。星形胶质细胞在凝胶中培养1、3、5、7 d,观察细胞活性以及细胞骨架的变化。结果:随着凝胶浓度的增加,琼脂糖凝胶弹性模量逐渐增加。2%的琼脂糖凝胶的弹性模量最接近筛板组织的弹性模量;在2%、3%琼脂糖凝胶环境下,细胞活性具有较高水平。随着细胞在凝胶中培养的时间增加,星形胶质细胞的突起逐渐伸出,细胞从球形向梭形或星形转变,更接近细胞真实的生长状态。结论:2%琼脂糖凝胶最接近星形胶质细胞在体内的生长环境,细胞成活率较高,是细胞体外三维培养的理想环境。

Objective To prepare different concentrations of agarose gels for the three-dimensional astrocytes culture in vitro, thereby finding the ideal environment for astrocytes culture in vitro. Methods The agarose gels at the mass concentrations of 1%, 2% and 3% were prepared and their elastic moduli were measured using nanoindentation test. The astrocytes were cultured in the gels for1, 3, 5 and 7 days to observe cell activity and cell cytoskeletal changes. Results The elastic moduli of agarose gels gradually increased with increasing gel concentration. The elastic modulus of 2% agarose gel was closest to that of the lamina cribrosa. The cell activity was higher in 2% and 3% agarose gels than 1% agarose gels. As the culture time increased, the processes of astrocytes gradually protruded, and the cells were changed from spherical to spindle or starlike, closer to the actual growth state of cells.Conclusion 2% agarose gel is closest to the in vivo environment of astrocytes, and the cell viability in 2% agarose gel is higher.Therefore, 2% agarose gel is an ideal environment for three-dimensional cell culture in vitro.