一种快速分离C_2C_(12)细胞核仁的方法研究  

A Quick New Method for Isolating the Nucleolus of C_2C_(12)

作　　者：邓恭华[1] 王慷慨[1] 刘可[1] 王浩[1] 易宇欣[1] 吴晓英[2] 肖献忠[1] 

机构地区：[1]中南大学湘雅医学院病理生理学教研室,休克研究室,湖南长沙410078 [2]中南大学湘雅医学院电镜室,湖南长沙410078

出　　处：《医学临床研究》2007年第4期529-531,共3页Journal of Clinical Research

基　　金：国家973重点项目(G2000056908);国家自然科学基金(30300177;30470851)资助

摘　　要：【目的】探讨一种快速分离核仁且能保持其完整结构与功能的方法。【方法】采用低渗裂解分离细胞核,密度梯度离心分离细胞核仁,电镜、免疫印迹检测核仁的纯度。【结果】电镜检测发现在放大2000倍时C2C12肌源细胞核仁呈现为大小基本一致、形态较为规则的黑色阴影,放大25000倍时可见单个网孔状的核仁,且核仁各组分清晰可见。免疫印迹检测发现核仁纯度较高,没有胞浆蛋白的污染。【结论】本研究建立了一种快速分离细胞核仁的方法。[Objective]To establish a rapid method for isolating nucleolus with intact structure and function. [Methods]The nucleoli were isolated by using hypotonic buffer and isolated and purified by Percoll liquids. Electronic-microscopy and immunoblotting were used to detect the quality of nucleoli. [Results]Nucleoli of C2 C12 myogenic cell were black granules with the same size under low-magnifying and the components of nucleoli could be seen clearly under high-magnifying electronic-microscope. Immunoblotting showed that the isolated nucleoli had high purity without contaminant derived from cytoplasmic proteins. [Conclusion]A rapid method for isolating nucleoli with intact structure and function is established.

关 键 词：核仁组成区 细胞分离/方法 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]