脑组织外泌体提取、鉴定及脑室注射活体示踪方法建立  

The establishment of brain tissue exosomes extraction, identification and intraventricular injection of vivo tracer

作　　者：王晋晨 叶樱泽 钟懿 古丽娟[2] 熊晓星[1] Wang Jinchen;Ye Yingze;Zhong Yi(Department of Neurosurgery,Renmin Hospital of Wuhan University,Wuhan 430060;不详)

机构地区：[1]武汉大学人民医院神经外科,430060 [2]武汉大学人民医院中心试验室,430060

出　　处：《卒中与神经疾病》2020年第1期9-13,共5页Stroke and Nervous Diseases

基　　金：国家自然科学基金资助项目(编号为81571147,81870939)。

摘　　要：目的探讨脑组织外泌体提取、鉴定及脑室注射活体示踪方法。方法采用蔗糖梯度离心法,多次离心去除杂质,最终得到纯度较高的脑组织外泌体,采用电镜及western-blot技术对不同密度蔗糖溶液内的外泌体进行鉴定,DiR (1,1’-dioctadecyltetramethyl indotricarbocyanine Iodide)染料标记后在近红外二区仪器下观察脑室注射外泌体后在体内的远程转移情况。结果应用蔗糖梯度离心法进行超高速离心可得到背景较为干净的脑组织外泌体,DiR染料可对外泌体进行标记并显示外泌体在体内转移途径。结论应用蔗糖梯度离心法可成功提取脑组织中外泌体,DiR染料可对外泌体标记并进行活体示踪。Objective To investigate the methods of brain tissue exosomes extraction, identification and intraventricular injection of vivo tracer.Methods The sucrose gradient centrifugation method was used to remove impurities and obtain higher purity brain tissue exosomes. The exosomes in different density sucrose solutions were identified by electron microscopy and western-blot.After DiR(1,1’-Dioctadecy ltetramethy lindotri carbocyanine Iodide) labeling, the exosomesdistant metastasis in the body after intraventricular injection was observed under near-infrared II imaging.Results sucrose gradient centrifugation could obtain brain tissue exosomes with a relatively clean background. DiR dye could label exosomes and show exosome distant metastasis in vivo. Conclusion Exosomes in brain tissue could be successfully extracted by sucrose gradient centrifugation. DiR dye could be labeled with exosomes and traced in vivo.

关 键 词：外泌体 蔗糖梯度离心法 脑室注射活体示踪 

分 类 号：R741.05[医药卫生—神经病学与精神病学]