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Published on: February 28, 2025
Harnessing 3D cultured MSC exosomes through tangential flow filtration for enhanced diabetic wound healing
Yanmei Chen1, Yang Xu2,3, Yali Zheng1
1Translational Research Centre of Regenerative Medicine and 3D Printing, Department of Orthopaedic Surgery, Guangzhou Key Laboratory of Spine Disease Prevention and Treatment, Guangdong Province Engineering Research Center for Biomedical Engineering, State Key Laboratory of Respiratory Disease, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, Guangdong, China.
Introduction:
Mesenchymal stem cell-derived exosomes have garnered considerable attention in regenerative medicine due to their non-immunogenicity, low infusion toxicity, easy accessibility, straightforward preservation, and minimal ethical concerns. While ultracentrifugation is the prevailing method for high-purity exosome isolation, it is limited by low throughput and the need for specialized infrastructure. This study investigates tangential flow filtration (TFF) as a promising alternative for exosome isolation. This technique offers simpler operation, higher yields, and improved recovery rates compared to ultracentrifugation.
Methods:
Human umbilical cord mesenchymal stem cells (hUCMSCs) were cultured in a 3D microcarrier-bioreactor system, and exosomes were extracted from the conditioned medium using either ultracentrifugation or an automated and enclosed TFF system. Subsequently, we compared the quantity, quality and therapeutic efficacy of the exosomes isolated via both approaches, evaluating their effects in vitro and in a mouse model of diabetic wound healing.
Results:
Our findings demonstrate that the TFF method effectively isolates high-quality exosomes that meet the standards set by the Minimum Information for Studies of Extracellular Vesicles (MISEV) 2023 guidelines, while achieving a significantly higher extraction yield compared to the traditional ultracentrifugation. Furthermore, both TFF and ultracentrifugation-derived exosomes demonstrate comparable biological activity in vitro and similar therapeutic potential for treating diabetic wound healing, potentially through promoting M2 macrophage polarization and angiogenesis.
Conclusion:
The results indicate that TFF is a viable method for scalable and efficient exosome production, facilitating advancements in clinical applications for diabetic wound repair.
