多聚左旋赖氨酸对大脑皮层星形胶质细胞体外培养的影响.docVIP

多聚左旋赖氨酸对大脑皮层星形胶质细胞体外培养的影响 　　[摘要] 目的 探讨不同浓度多聚左旋赖氨酸（PLL）对体外培养大脑皮层星形胶质细胞（AS）的影响。 方法 采用酶消法、差速贴壁获得大脑皮层AS细胞，细胞分为四组，分别种植于对照组（未包被PLL）、25 μg/mL PLL包被组、50 μg/mL PLL包被组、100 μg/mL PLL包被组包被的培养瓶或板内，运用细胞计数检测细胞贴壁率及CCK检测法绘制原代细胞生长曲线；待细胞铺满瓶底，予振荡及传代法去除其他类型细胞。第三代AS，通过胶质纤维酸性蛋白（GFAP）免疫荧光计数细胞纯度及CCK检测细胞活性。 结果 细胞贴壁率随PLL包被浓度的增加而增加；原代AS生长曲线显示对照组生长速度较三组PLL包被组慢；除100 μg/mL PLL包被组[（90±0.53）%]外，其余各组细胞纯化率均大于95%；对照组第4、6天细胞活性均明显低于各PLL包被组，差异均有高度统计学意义（P 0.01）。 结论 PLL能有效地促进AS的贴壁和生长，适宜包被浓度为25～50 μg/mL。 　　[关键词] 星型胶质细胞；体外培养；多聚左旋赖氨酸；免疫荧光；胶质纤维酸性蛋白 　　[中图分类号] Q421 [文献标识码] A [文章编号] 1673-7210（2016）05（a）-0004-05 　　[Abstract] Objective To investigate the effects of different concentrations of poly-L-lysine （PLL） on the cultures of cerebra lcorticalastrocytes （AS） in vitro. Methods Astrocytes were dissociated from cerebra lcortices with the methods of trypsinization and differential adherence. They were divided into four groups and respectively planted in culture flasks or plates which coated PLL of different concentrations： control group （non-PLL）， 25 μg/mL PLL envelop group， 50 μg/mL PLL envelop group， 100 μg/mL PLL envelop group. Adherent rates were detected by counted number and cell growth curves were drawn with CCK-8 colorimeter. To remove other cell types， the cells were shaken and passaged when they were confluent in a monolay on the bottom. Purities were assessed by immune of luorescence analysis of glial fibrillate acidic protein （GFAP） and cell viabilities were measured by CCK-8 colorimeter in the third generation astrocytes. Results Adherent rates were increased in a dose-dependent manner. Growth curves showed that the growth speed of control group was significantly slower than three PLL envelop groups. The purities of all groups were over 95% except 100 μg/mL PLL envelop group [（90.00±0.53）%]. The cell activities of control group on the fourth day and the sixth day both were lower than three PLL envelop groups， the differences were statistially signifiant （P 0.01）. Conclusion PLL-coated techni