دورية أكاديمية
Development of an optimized and scalable method for isolation of umbilical cord blood‐derived small extracellular vesicles for future clinical use
| العنوان: | Development of an optimized and scalable method for isolation of umbilical cord blood‐derived small extracellular vesicles for future clinical use |
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| المؤلفون: | Renato M. S. Cardoso, Silvia C. Rodrigues, Claudia F. Gomes, Filipe V. Duarte, Maryse Romao, Ermelindo C. Leal, Patricia C. Freire, Ricardo Neves, Joana Simões‐Correia |
| المصدر: | Stem Cells Translational Medicine, Vol 10, Iss 6, Pp 910-921 (2021) |
| بيانات النشر: | Oxford University Press, 2021. |
| سنة النشر: | 2021 |
| المجموعة: | LCC:Medicine (General) LCC:Cytology |
| مصطلحات موضوعية: | angiogenesis, cell signaling, cellular therapy, clinical translation, microRNA, stem cells, Medicine (General), R5-920, Cytology, QH573-671 |
| الوصف: | Abstract Extracellular vesicles (EV) are a promising therapeutic tool in regenerative medicine. These particles were shown to accelerate wound healing, through delivery of regenerative mediators, such as microRNAs. Herein we describe an optimized and upscalable process for the isolation of EV smaller than 200 nm (sEV), secreted by umbilical cord blood mononuclear cells (UCB‐MNC) under ischemic conditions and propose quality control thresholds for the isolated vesicles, based on the thorough characterization of their protein, lipid and RNA content. Ultrafiltration and size exclusion chromatography (UF/SEC) optimized methodology proved superior to traditional ultracentrifugation (UC), regarding production time, standardization, scalability, and vesicle yield. Using UF/SEC, we were able to recover approximately 400 times more sEV per mL of media than with UC, and upscaling this process further increases EV yield by about 3‐fold. UF/SEC‐isolated sEV display many of the sEV/exosomes classical markers and are enriched in molecules with anti‐inflammatory and regenerative capacity, such as hemopexin and miR‐150. Accordingly, treatment with sEV promotes angiogenesis and extracellular matrix remodeling, in vitro. In vivo, UCB‐MNC‐sEV significantly accelerate skin regeneration in a mouse model of delayed wound healing. The proposed isolation protocol constitutes a significant improvement compared to UC, the gold‐standard in the field. Isolated sEV maintain their regenerative properties, whereas downstream contaminants are minimized. The use of UF/SEC allows for the standardization and upscalability required for mass production of sEV to be used in a clinical setting. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 2157-6580 2157-6564 |
| العلاقة: | https://doaj.org/toc/2157-6564Test; https://doaj.org/toc/2157-6580Test |
| DOI: | 10.1002/sctm.20-0376 |
| الوصول الحر: | https://doaj.org/article/e1f2c32bd9914a6886fa4ef15b781b83Test |
| رقم الانضمام: | edsdoj.1f2c32bd9914a6886fa4ef15b781b83 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 21576580 21576564 |
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| DOI: | 10.1002/sctm.20-0376 |