6.1.2 外泌体示踪-体外示踪-慢病毒载体构建GFP的稳定细胞系

分子运输在真核细胞中是一个动态的过程，Cyto-Tracers™提供了照亮细胞隔间以监测细胞器的运动和定位，并追踪内吞作用和胞吐作用的能力。SBI已经创建了一系列基于慢载体的Cyto-Tracers™，利用GFP融合蛋白标记细胞隔间、细胞器、囊泡和结构，使更长期和更深入的实验。Cyto-Tracers可用于转染，也可包装成病毒，在原代细胞、肿瘤细胞系和干细胞中创建稳定的GFP示踪细胞系。SBI 提供商品化的已经融合外泌体表面marker的Cyto-Tracers 系列产品，用于构建稳定表达荧光的细胞系。

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pCT-CD63-GFP (pCMV, Exosome/Secretory, CD63 Tetraspanin Tag)
特点
基于entivector 的稳定系统
共定位研究的理想选择
实时监测外泌体动态和功能研究
标记原代细胞、肿瘤细胞、干细胞等的外泌体

图 9 From Garcia, et al. Exosome transfer from CMs to ECs. 
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(A) H9c2 transfected with pCT-CD63-GFP. FACS analysis of 90% GFP positive cells.
 (B) Representative images from confocal time-lapse microscopy of mouse ECs (ACTB-DsRed) (EC; red) co-cultured with H9C2-CD63-GFP cells (green), previously cultured for 24 h in +/-St medium. Exosome transfer from H9C2 CMs to EC can be observed (S1 and S2 Movies, below). White arrows show CD63-GFP structures inside ECs
 (C) Representative immunostaining of H9C2-CD63-GFP and HUVEC co-cultures; anti-GFP (green) and anti-CD31 (red). The images illustrate GFP fluorescence from CD63-GFP exosomes in CD31-positive cells (red) after 24 h incubation in +/-St medium. White arrows show CD63-GFP structures inside ECs.

视频链接： https://www.systembio.com/exosome-cyto-tracer-pct-cd63-gfp