人肺癌A549细胞的侧群细胞流式分选方法优化.doc

文档介绍：
人肺癌 A549 细胞的侧群细胞流式分选方法
优化#
陈洋,杨丽娟,王通**
5
10
(暨南大学生命与健康工程研究院,功能蛋白质研究广东普通高校重点实验室,
广州 510632)
摘要:具外排 Hochest 33342 染料能力的侧群(side population, SP)细胞有癌症干细胞(cancer
stem cell, CSC)样特征,可能富含一定比例的 CSC。然而传统流式分选方法只能得到少量
侧群细胞。本研究利用流式细胞术分析发现,侧群细胞中反映细胞颗粒度的参数 SSC (side
scatter, SSC)较主群细胞小,利用这一特征划门排除 SSC 较大的细胞有利于提高侧群细胞检
测分辨率,达到提高分选效率和分选纯度的目的。本研究建立了新的侧群细胞分选划门策略,
为 CSC 研究提供优化的实验方法。
关键词:侧群细胞;流式细胞分选;癌症干细胞
中图分类号:Q2-33
15
Optimization of flow cytometric sorting on the side
population of human lung A549 cells
Chen Yang, Yang Lijuan, Wang Tong
(Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes,
20
25
30
35
40
Institute of Life and Health Engineering, College of Life Science and Technology, Jinan
University, GuangZhou 510632)
Abstract: Side population (SP) cells are able to efflux Hoechst 33342 dye. They have cancer stem
cell (CSC) -like features, suggesting an enrichment of CSC. However, the traditional flow
cytometric (FACS) methods for SP sorting have low acquisition efficiency. To solve this
hindrance, we demonstrated in this study that the parameter of side scatter (SSC), an indicate of
granularity, of SP cells are considerably smaller than the cells of main population (MP). We
therefore excluded larger cells in the MP cells and acquired high purity side population cells with
much improved sorting efficiency. We established a