首页|UC法和exoEasy法提取鹿茸间充质干细胞外泌体的比较

UC法和exoEasy法提取鹿茸间充质干细胞外泌体的比较

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经原代体外分离培养及鉴定获取鹿茸间充质干细胞(Antler mesenchymal stem cells,AMSCs),分别利用超速离心法(Ultracentrifugation,UC)和膜亲和法(exoEasy)提取AMSCs培养基上清中的外泌体(Exosomes,Exo),并比较2种方法提取AMSCs-Exo效果,以及验证AMSCs-Exo的功能。结果表明:UC法和exoEasy法均能够提取到AMSCs-Exo,UC法提取的AMSCs-Exo结构更完整,杂质较少,粒径分布具有代表性,仅有单一主峰,Western blot(WB)能检测出AMSCs-Exo标志蛋白;而exoEasy法提取的AMSCs-Exo浓度较低,结构不完整,杂质较多,并且颗粒分布分散,不具有代表性,没有单一主峰,WB无法检测到Exo标志蛋白。进一步通过5-乙炔基-2'脱氧尿嘧啶核苷(5-ethynyl-2'-deoxyuridine assay,EdU)试验证实,AMSCs-Exo可显著促进haCat细胞增殖。通过比较2种方法提取的AMSCs-Exo可知,UC法提取的AMSCs-Exo更适合相关功能验证研究中。
Comparison between UC Method and ExoEasy Method for Extract-ing Exosomes from Antler Mesenchymal Stem Cells
After primary isolation and identification of antler mesenchymal stem cells(AMSCs),exo-somes were isolated from AMSCs culture medium by UC(Ultracentrifugation)and exoEasy methods.The effects of AMSCs-Exo extracted with two methods were compared,and the functions of AMSCs-Exo were verified.The results showed that AMSCs-Exo could be isolated by both UC method and exoEasy method.The isolated AMSCs-Exo by UC method had more complete structure,fewer impuri-ties,and more representative particle size distribution,compared with AMSCs-Exo isolated by exoEasy method.It had a single main peak and AMSCs-Exo protein marker could be detected by WB(Western blot).The isolated AMSCs-Exo by exoEasy method had lower concentration,more incom-plete structure,more impurities,more dispersed particles,and more unrepresentative distribution,compared with AMSCs-Exo isolated by UC method.It had no single main peak and no detectable pro-tein marker.EdU results showed that AMSCs-Exo could promote the proliferation of haCat cells sig-nificantly.By comparing the two methods,UC method is more suitable for relevant functional verifi-cation studies.

antlermesenchymal stem cellexosomeUC methodexoEasy method

张琦、杨春、孙胜楠、郭佳、路洪涛、刘莹、杨敏、彭英华

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中国农业科学院特产研究所,长春 130112

鹿茸 间充质干细胞 外泌体 超速离心法 膜亲和法

吉林省科技发展计划技术攻关项目中国农业科学院科技创新工程项目

20190301082NYCAAS-ASTIP-2019-ISAPS

2024

吉林农业大学学报
吉林农业大学

吉林农业大学学报

CSTPCD北大核心
影响因子:1.014
ISSN:1000-5684
年,卷(期):2024.46(1)
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