Li Jing, Shen Yi, Tang Wei-xue, Chen Li, Duan Hong
Department of Pathophysiology, College of Basic Medicine, Chongqing University of Medical Sciences, Chongqing 400016, China.
Zhonghua Gan Zang Bing Za Zhi. 2007 Jun;15(6):437-40.
To separate and identify the exosomes derived from a mouse hepatoma carcinoma cell line (H22) and to detect their protein composition, and to investigate the possibility of using these exosomes as a kind of tumor vaccine.
Exosomes were purified by serial ultracentrifugation and sugar density ultracentrifugation, and then they were observed and identified by electron microscopy. Exosomes underwent peptide mass fingerprint and Western blot analyses.
H22 cell-derived exosomes were 20-90 nm round or oval vesicles. The exosomes expressed HSP70, ICAM-1, EF-G2, DLC-A, C-myc protein and Vav-2 protein.
Serial ultracentrifugation and sugar density ultracentrifugation can be used to purify H22 cell-derived exosomes. H22 cell-derived exosomes express a distinct set of proteins involving in and/or relating to antigen presentation (HSP70, ICAM-1), migration (DLC-A), adhesion (ICAM-1), cytoskeleton (EF-G2) and tumour antigens (C-myc, Vav-2). The exosomes have immunogenicity.
分离并鉴定源自小鼠肝癌细胞系(H22)的外泌体,检测其蛋白质组成,并研究将这些外泌体用作一种肿瘤疫苗的可能性。
通过连续超速离心和蔗糖密度超速离心纯化外泌体,然后用电子显微镜进行观察和鉴定。对外泌体进行肽质量指纹图谱分析和蛋白质印迹分析。
H22细胞来源的外泌体为20 - 90纳米的圆形或椭圆形囊泡。这些外泌体表达热休克蛋白70(HSP70)、细胞间黏附分子1(ICAM-1)、延伸因子G2(EF-G2)、DLC-A、C-myc蛋白和Vav-2蛋白。
连续超速离心和蔗糖密度超速离心可用于纯化H22细胞来源的外泌体。H22细胞来源的外泌体表达一组独特的蛋白质,这些蛋白质参与和/或与抗原呈递(HSP70、ICAM-1)、迁移(DLC-A)、黏附(ICAM-1)、细胞骨架(EF-G2)及肿瘤抗原(C-myc、Vav-2)相关。这些外泌体具有免疫原性。