Preclinical Characterization and In Vivo Imaging of ¹¹¹In-Labeled Mesenchymal Stem Cell–Derived Extracellular Vesicles

Purpose: Mesenchymal stem cell–derived EVs (MSC-EVs) are demonstrated to have similar therapeutic effect as their cells of origin and represent an attractive cell-free stem cell therapy. With the potential to be the future medical regimen, the information of fate and behavior of MSC-EVs in the living subject should be urgently gathered. This study aimed to track MSC-EVs by ¹¹¹In-labeling and μSPECT/CT imaging. Procedures: Wharton’s jelly-MSC-EVs (WJ-MSC-EVs) were isolated using Exo-Prep kit followed by characterization of expressing markers and size. After labeled by ¹¹¹In-oxine, ¹¹¹In-EVs were injected into C57BL/6 mice followed by μSPECT/CT imaging. Organs were then taken out for ex vivo biodistribution analysis. Results: The radiochemical purity of ¹¹¹In-EVs was > 90 % and remained stable up to 24 h. The image results showed that with injection of ¹¹¹In-EVs, the signal mainly accumulated in the liver, spleen, and kidney, compared to that in lung and kidney after ¹¹¹In-oxine injection. The ex vivo biodistribution showed the similar pattern to that of imaging. Chelation of free ¹¹¹In with EDTA was found necessary to reduce the nonspecific accumulation of signal. Conclusion: This study demonstrated the feasibility of radiolabeling WJ-MSC-EVs with ¹¹¹In-oxine for in vivo imaging and quantitative analysis in a mouse model. This simple and quick labeling method preserves the characteristics of WJ-MSC-EVs. The results in this study provide a thorough and objective basis for future clinical study.