诺考达唑和阿非科林对人胚胎干细胞周期的影响  

Effects of nocodazole and aphidicholine to cell cycle of human embryonic stem cells

作　　者：魏建峰[1] 王昱博[2] 

机构地区：[1]徐州医学院基础医学院组织胚胎学教研室,江苏徐州221004 [2]吉林大学临床医学院,吉林长春130021

出　　处：《生物技术》2015年第6期591-596,共6页Biotechnology

基　　金：江苏省高校自然科学基金项目("microRNA对干细胞神经谱系发育影响的初步探究";No.14KJB310021);江苏省脑病生物信息重点实验室开放课题("microRNAs在神经谱系分化命运决定中的作用机制研究";No.JSBl1403)资助

摘　　要：[目的]探究诺考达唑和阿非科林联合应用对人胚胎干细胞周期同步化到G1和S期的作用并观察药物对细胞生长和生物功能的影响。[方法]采用诺考达唑和阿非科林先后作用于胚胎干细胞,流式细胞仪测定细胞周期,显微镜下观察药物对细胞生长状态的影响,荧光染色技术观察胚层标志物α-SMA和β-tubulin表达。[结果]加入诺考达唑和阿非科林后,G1期为66.35%,S期为33.52%,多数细胞被同步化在G1期;诺考达唑作用16 h,G1期为18.45%,S期为79.10%,多数细胞从周期抑制中恢复;能够形成典型的拟胚体;荧光染色显示α-SMA和β-tubulin均有表达。[结论]诺考达唑和阿非科林联合作用能够使胚胎干细胞66.35%同步化在G1期,诺考达唑作用16 h,79.10%的细胞同步化在S期。中胚层标志物α-平滑肌肌动蛋白和外胚层标志物β微管蛋白在蛋白水平均有明显表达。去除药物后,细胞未发生显著性损伤。[ Objective] To investigate the effects of nocodazole and aphidicholine to cell cycle when embryonic stem cell cycle synchronization to G1 and S phase,and observe the change of cell growth and function after medicine were used. [ Methods] Nocodazole and aphidicholine were given to human embryonic stem cells, cell cycle was measured by flow cytometry, the effects of drug for cell growth was detected by light microscope, and the expression of germ layer markers α - SMA and β- tubulin were analyzed by immunofluorescence staining. [ Results ] After addition of nocodazole and aphidicholine, embryonic stem cells can be synchronized to corresponding cell cycle, G1 phase was 66.35%, S phase is 33.52%, the majority of the cells were syn- chronized in G1 phase, cell growth state can be restore after removal of the drug, and embryonic stem cells have the ability to form embryoid bodies and express α - SMA and β- tubulin by immunofluorescence. [ Conclusion I Nocodazole and the com- bined effects of aphidicholine, enabling 66.35% of embryonic stem cells synchronized in G1 phase, nocodazole effect 16 h, 79.10% of the cells synchronized in S phase. Mesodermal markers α - smooth muscle actin and ectoderm markers β- tubulin express significantly. After removal of the drug, the cells no significant damage occurred.

关 键 词：胚胎干细胞 诺考达唑 阿非科林 细胞周期 

分 类 号：R965.2[医药卫生—药理学]