根尖牙乳头干细胞外泌体的提取与鉴定  

Isolation and identification of exosomes derived from stem cells from apical papilla

作　　者：余思 蒋欢 刘尧[1] 戴璐 陈旭[1] YU Si;JIANG Huan;LIU Yao;DAI Lu;CHEN Xu(Department of Pediatric Dentistry,School and Hospital of Stomatology,China Medical University,Shenyang 110002,China)

机构地区：[1]中国医科大学口腔医学院.附属口腔医院儿童口腔科,辽宁沈阳110002

出　　处：《中国实用口腔科杂志》2018年第6期347-350,共4页Chinese Journal of Practical Stomatology

基　　金：国家自然科学基金面上项目(81771059);军事口腔医学国家重点实验室开放课题(2017KA04);辽宁省自然科学基金项目项目(201602842)

摘　　要：目的提取根尖牙乳头干细胞(stem cells from apical papilla,SCAP)来源的外泌体并对其鉴定。方法原代培养SCAP并鉴定,收集SCAP培养上清,采用超速离心法提取其外泌体,透射电镜下观察外泌体形态,免疫印记法检测外泌体特异性标记蛋白CD9、Alix的表达。结果 SCAP能分泌外泌体,电镜下可见其为呈杯状的囊泡结构,直径30~100 nm;特异性标记蛋白CD9、Alix呈阳性表达。结论采用超速离心法可以成功从SCAP培养上清中提取外泌体。Objective To isolate and identify exosomes derived from stem cells from apical papilla. Methods Stem cells from apical papilla（SCAP）were primarily cultured and identified. SCAP-derived exosomes were isolated by ultracentrifugation. The morphology was observed by transmission electron microscopy. The markers CD9 and Alix were detected by Western blotting. Results SCAP could release exosomes,which showed a cup-like microvesicle structure,with a diameter of about 30-100 nm under electronic microscopy. The specific markers CD9 and Alix were positively expressed. Conclusion The results show that exosomes can be successfully isolated from culture supernatants of SCAP by ultracentrifugation.

关 键 词：根尖牙乳头干细胞 外泌体 超速离心 特异性标记物 

分 类 号：R78[医药卫生—口腔医学]