张鹏,舒莉萍,周艳华,范安然
ZHANG Peng,SHU Liping,ZHOU Yanghua,FAN Anran

• 1:贵州医科大学细胞工程生物医药技术国家地方联合工程实验室组织工程与干细胞实验中心贵州省再生医学重点实验室
• 2:中国医学科学院成体干细胞转化研究重点实验室

摘要(Abstract)：

目的:验证HEK293-Tau细胞的外泌体中是否存在Tau蛋白、及Tau蛋白是否能够在细胞间通过外泌体传播。方法:通过经典的超速离心结合超滤法分离HEK293-Tau细胞的外泌体,通过透射电镜分析外泌体形态、采用Western blot鉴定外泌体特异性标记物(HSP70、CD63及CD9)及外泌体和细胞培养液中Tau蛋白表达。结果:超速离心结合超滤法成功地分离了HEK293-Tau细胞的外泌体,在透射电镜下观察到外泌体呈现典型的杯状形态,分离的外泌体中检测到HSP70、CD63及CD9等外泌体特异性标记物的表达;在外泌体及细胞培养液中能够检测到Tau蛋白的存在。结论:HEK293-Tau细胞外泌体中存在Tau蛋白,并且Tau蛋白可以通过外泌体传递到细胞外空间。
Objective: To examine whether Tau protein exists in exosomes isolated from supernatants derived from culture media with HEK293-Tau and whether the Tau protein can be delivered into extracellular space via exosomes. Methods: Exosomes were isolated using ultracentrifugation combined with ultrafiltration. The morphology of isolated exosomes were observed by a transmission electron microscopy. The marker of exosomes was further confirmed using Western blot. Tau protein in isolated exosomes and cell culture medium was also checked using Western blot. Results: Isolated exosomes showed typically cup-shaped morphology. In addition, the exosome marker HSP70, CD63 and CD9 and Tau protein are able to detect in exosomes and cell culture medium. Conclusions: Tau present in cell culture media might be delivered by exosomes derived from HEK293-Tau cells.

关键词(KeyWords)： Tau蛋白;外泌体;阿尔茨海默综合征;HEK293-Tau细胞;神经退行性疾病;超速离心法;超滤法
Tau protein;exosomes;Alzheimer's disease;HEK293-Tau cell;neurodegenerative disease;ultracentrifugation;ultrafiltration

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(31660316);; 中国医学科学院中央级公益性科研院所基本科研业务费专项资金资助(2018PT31048);; 贵州省应用基础研究计划重大专项“肿瘤转移复发耐药机制的研究及临床治疗新技术”[黔科合J重大字(2015)2003]

作者(Author): 张鹏,舒莉萍,周艳华,范安然
ZHANG Peng,SHU Liping,ZHOU Yanghua,FAN Anran

DOI: 10.19367/j.cnki.1000-2707.2019.06.003

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