DSPE-PEG-SH与DSPE-PEG-FITC在EV修饰中的对比研究-UDP糖丨MOF丨金属有机框架丨聚集诱导发光丨荧光标记推荐西安齐岳生物

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DSPE-PEG-SH与DSPE-PEG-FITC在EV修饰中的对比研究

发布时间：2025-07-07 作者：zyl 分享到：

文献：Extracellular Vesicle Crosslinkers Constructing Hydrogels with Stress-Relaxation and Bioactive Protein Modification

作者：Lufeng Shi, Yanzhen Jing, Haowen Lu, Fanxuan Zhao, Mengying An, Shuiling Jin, Chang Gao, Yongdong Dai, Yinxin Zhu, Shuxu Yang, Songying Zhang, Xuesong Ye, Xiujun Cai … See all authors

文献链接：https://advanced.onlinelibrary.wiley.com/doi/full/10.1002/admi.202400885

We began by using the amphiphilic molecule, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(polyethylene glycol) (DSPE-PEG), with functional groups on the PEG side to incubate with EVs secreted by stem cells from human exfoliated deciduous teeth (SHEDs). This approach allowed the amphiphilic molecules to insert into the EV bilayer membrane, displaying functional groups on the EV surface (Figure 1a). The EVs were dispersed in aqueous solution at a concentration of 1.8 × 10¹⁰ EVs mL−1, with an average size of 151.4 nm (Figure S1, Supporting Information). To confirm the successful insertion of the amphiphilic molecules into the EV membrane, we employed DSPE-PEG-thiol (DSPE-PEG-SH) and DSPE-PEG-fluorescein isothiocyanate (DSPE-PEG-FITC) as examples for this modification process. Flow cytometry analysis revealed a significant increase in mean fluorescent intensity (MFI) of EVs after incubation with DSPE-PEG-FITC compared to controls without incubation or with a 0.5-h incubation of DSPE-PEG-SH, confirming the successful insertion of amphiphilic molecules into the EV membrane (Figure 1b). Additionally, the comparison between EVs incubated with DSPE-PEG-FITC and DSPE-PEG-SH further validated the accuracy of our gating strategy in flow cytometry.

DSPE-PEG-FITC

我们首先使用两亲性分子，1,2-二硬脂酰基-sn-甘油-3-磷酸乙醇胺-N-（聚乙二醇）（DSPE-PEG），在PEG侧具有官能团，与人类脱落乳牙（SHED）干细胞分泌的EV一起孵育。这种方法允许两亲性分子插入EV双层膜中，在EV表面显示官能团。

EV以1.8×10¹的浓度分散在水溶液中⁰ EV mL−1，平均粒径为151.4 nm（图S1，支持信息）。为了确认两亲性分子成功插入EV膜中，我们采用了DSPE-PEG-SH和DSPE-PEG-异硫氰酸荧光素（DSPE-PEG-FITC）作为这种修饰过程的例子。

流式细胞术分析显示，与未孵育或孵育0.5小时的DSPE-PEG-SH的对照组相比，与DSPE-PEG-FITC孵育后，EV的平均荧光强度（MFI）显著增加，证实了两亲性分子成功插入EV膜。此外，用DSPE-PEG-FITC和DSPE-PEG-SH孵育的EV之间的比较进一步验证了我们在流式细胞术中门控策略的准确性。