外泌体不同制备方法的综合实验教学实践  

Comprehensive Experimental Teaching Practice on Various Exosome Isolation Methods

作　　者：李艳伟[1] 郭春 周南 黄琼[1] 王佳佳[1] 陈静瑶[1] 黄莹莹[1] 沈鑫 于晓敏 沈红英 LI Yanwei;GUO Chun;ZHOU Nan;HUANG Qiong;WANG Jiajia;CHEN Jingyao;HUANG Yingying;SHEN Xin;YU Xiaomin;SHEN Hongying(Core Facilities,School of Medicine Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学医学院公共技术平台,杭州310058

出　　处：《中国细胞生物学学报》2024年第6期1191-1198,共8页Chinese Journal of Cell Biology

基　　金：浙江省自然科学基金分析测试项目(批准号:LTGC23C070003)资助的课题。

摘　　要：流式细胞术随着单克隆抗体技术和光电技术的快速发展而日趋多元化和多样化。然而,该技术仍受流式细胞仪检测极限的限制而制约外泌体等细胞外囊泡的分离纯化。该文通过对外泌体样本制备方法的建立和探索,使流式细胞技术理论与实验教学内容更加直观、更易理解和掌握。该方法包括对流式细胞仪配置及实验参数的优化、外泌体的不同制备方法(超速离心、流式细胞分选)、纳米流式分析鉴定、电镜成像鉴定等步骤。通过对该方法的实践操作,使学生充分掌握相关的检测技术和数据分析手段,为流式细胞术在外泌体分选及后续功能研究中提供支撑作用。教学、科研、实践相融合,极大地激发了学生学习实验技能的兴趣和积极性,大大提升了学生实验技能水平以及科研创新能力等科研素养,为新时期全能型科研创新人才的培养提供助力。FCM(flow cytometry)has become increasingly diverse and versatile with the rapid development of monoclonal antibody technology and optoelectronic technology.However,the isolation and purification of exosomes and other extracellular vesicles have been impeded by the limitations of the detection capabilities of FCM.This study establishes and investigates methods for optimizing exosome sample preparation to enhance the theoretical and practical aspects of FCM teaching,rendering it more intuitive,comprehensible,and feasible.The methods include optimizing FCM configuration and experimental parameters,different exosome isolation methods[ultracentrifugation,FACS(flow cytometric sorting),nanoFCM analysis,and electron microscopy imaging identification].Through practical application of these methods,students can fully master the relevant detection techniques and data analysis methods,providing support for the application of FCM in exosome isolation and subsequent functional studies.The integration of teaching,research,and practical experience greatly stimulates students’interest and enthusiasm for learning experimental skills,significantly enhancing their experimental skills and research innovation capabilities,contributing to the cultivation of versatile research and innovation talents in the new era.

关 键 词：外泌体 流式细胞术 超速离心 综合实验教学 

分 类 号：G642.423[文化科学—高等教育学] Q2-4[文化科学—教育学]