Journal Article

Extracellular vesicle manufacture via FACTORY: fully automated collection technology and optimum machinery for clinical translational applications

Chen, Li-Guo, Huang, Jue, Chen, Gang, Wu, Lian-Zhi, Xu, Jing-Yi, Liu, Xing-Chi, Zhang, Zhi-Ling, Yu, Zi-Li, Tian, Jia-Wen, Ye, Zi-Wu

2025

Overview

The large-scale automated manufacture of extracellular vesicles (EVs) is crucial for meeting the growing demand for EV-based therapies, enabling the standardized, efficient, and scalable production of EVs for clinical translation.We developed a fully automated collection technology and optimized machinery (FACTORY), achieving large-scale automated manufacture of EVs.The EVs manufactured via FACTORY maintain normal biological activity while meeting stringent quality standards: sterility, mycoplasma-free status, low endotoxin levels, and high consistency.The FACTORY platform allows researchers across disciplines to obtain EVs through a one-click operation, ensuring batch-to-batch consistency in isolation technology and comparability of research outcomes, thereby accelerating the clinical translation of EV-based applications. Extracellular vesicles (EVs) hold significant potential as therapeutic agents and drug carriers. However, current isolation techniques severely limit their clinical application, due to heavy reliance on manual operation, making large-scale isolation of EVs impractical and failing to meet the requirements for clinical translation. Here, we set up the fully automated collection technology and optimum machinery (FACTORY) platform, allowing the efficient collection of high-quality EVs. The platform integrates continuous flow centrifugation and tangential flow filtration (TFF) technologies, achieving a seamless process for the removal of impurities and collection of EVs, thereby ensuring that large scale-manufactured EVs are sterile, mycoplasma free, and low in endotoxins, and exhibit good consistency. We successfully obtained a substantial quantity of EVs utilizing FACTORY, and systematically characterized their EV-specific markers, biological functions, and therapeutic effects. Results indicated that FACTORY significantly promotes the clinical translation of EVs, thereby laying a solid foundation for their application in drug delivery and beyond. [Display omitted] The FACTORY platform addresses the urgent need for scalable, automated, and efficient methods for collecting EVs, which hold great promise in clinical therapeutics. Currently, FACTORY is assessed at Technology Readiness Level (TRL) 6, indicating its readiness for real-world implementation. By integrating continuous flow centrifugation and tangential flow filtration (TFF), FACTORY enables large-scale EV isolation with high yield, sterility, and batch-to-batch consistency. Experimental results show that FACTORY produces a higher yield of EVs compared with traditional ultracentrifugation, with comparable purity and morphology. In addition, FACTORY-isolated EVs meet critical safety criteria, including sterility, mycoplasma-free status, and minimal endotoxin levels. The FACTORY platform holds significant promise for advancing EV-based therapeutics, with potential applications in regenerative medicine, drug delivery, and immunomodulation. Its scalability, automation, and ability to produce high-quality EVs position it as a transformative tool for clinical and industrial use, paving the way for broader adoption in emerging medical fields, including medical aesthetics and targeted therapy development. This study developed the fully automated collection technology and optimum machinery (FACTORY) platform for extracellular vesicles (EVs), integrating continuous flow centrifugation and tangential flow filtration. The platform allows one-click, large-scale isolation of high-quality EVs with exceptional consistency, accelerating the clinical translation of EV-based applications.

• Publisher: Elsevier Ltd,Elsevier Limited
• Subject: automated platform / Automation / Biomarkers / biotechnology / Cell culture / Centrifugation / Centrifugation - methods / Clinical trials / Continuous flow / Current carriers / Drug carriers / Drug delivery / drugs / Efficiency / Endotoxins / EV isolation / Extracellular vesicles / Extracellular Vesicles - chemistry / Extracellular Vesicles - metabolism / filtration / Filtration - methods / Humans / Impurities / Internal Medicine / large-scale manufacture / manufacturing / Pharmacology / Pore size / Proteins / Regenerative medicine / Stem cells / therapeutics / Titanium alloys / Translation / translational applications / Translational Research, Biomedical / Wound healing