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Updated: Aug 4, 2025
Large-Scale Preparation of Synovial Fluid Mesenchymal Stem Cell-Derived Exosomes by 3D Bioreactor Culture
Published on: July 26, 2022
Three-dimensional-cultured MSC-derived exosome with hydrogel for cerebral ischemia repair
Min Han1, Zihao Zhang2, Zihao Liu3
1Department of Neurosurgery, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Jinan 250014, PR China; Medical Science and Technology Innovation Center, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan 250117, PR China.
Abstract:
Microglia-mediated neuroinflammatory response, one of the most essential pathological processes of cerebral ischemia-reperfusion (I/R) injury, is acknowledged as the main factors leading to poor prognosis of cerebral ischemia. Exosome derived from mesenchymal stem cell (MSC-Exo) exhibits neuroprotective functions by reducing cerebral ischemia-induced neuroinflammatory response and promoting angiogenesis. However, MSC-Exo has disadvantages such as insufficient targeting capability and low production, which limits their clinical applications. Here, we fabricated gelatin methacryloyl (GelMA) hydrogel for three-dimensional (3D) culture of MSCs. It is indicated that 3D environment could simulate the biological niches of MSCs, thereby significantly increasing the cell stemness of MSCs and improving the yield of MSCs-derived exosomes (3D-Exo). In this study, we utilized the modified Longa method to induce middle cerebral artery occlusion (MCAO) model. Additionally, in vitro and in vivo studies were conducted to interrogate the mechanism of the stronger neuroprotective effect of 3D-Exo. Furthermore, the administration of 3D-Exo in MCAO model could promote neovascularization in infarct region and result in a significant suppression of inflammatory response. This study proposed an exosome-based targeting delivery system for cerebral ischemia and provided a promising strategy for efficient and large-scale production of MSC-Exo.
Insights
Gelatin methacryloyl hydrogels enhance mesenchymal stem cell exosome production, offering a promising strategy to combat neuroinflammation and promote recovery after cerebral ischemia-reperfusion injury.
Area of Science:
- Biomedical Engineering
- Neuroscience
- Regenerative Medicine
Background:
- Microglia-driven neuroinflammation is a key factor in poor outcomes following cerebral ischemia-reperfusion (I/R) injury.
- Mesenchymal stem cell-derived exosomes (MSC-Exo) show neuroprotective potential but face limitations in targeting and production yield.
- Developing strategies for enhanced exosome production and delivery is crucial for treating cerebral ischemia.
Purpose of the Study:
- To develop a 3D culture system using GelMA hydrogels to improve MSC-Exo yield and efficacy.
- To investigate the neuroprotective mechanisms of exosomes derived from MSCs cultured in 3D (3D-Exo).
- To evaluate the therapeutic potential of 3D-Exo in a middle cerebral artery occlusion (MCAO) model of cerebral ischemia.
Main Methods:
- Fabrication of GelMA hydrogels for 3D culture of MSCs.
- Induction of cerebral ischemia-reperfusion injury using the modified Longa method (MCAO model).
- In vitro and in vivo assessments of 3D-Exo's effects on neuroinflammation, angiogenesis, and neurological function.
Main Results:
- 3D culture significantly enhanced MSC stemness and increased the yield of exosomes (3D-Exo).
- 3D-Exo demonstrated superior neuroprotective effects compared to conventional MSC-Exo.
- Administration of 3D-Exo in the MCAO model promoted neovascularization and suppressed inflammatory responses in the infarct region.
Conclusions:
- GelMA hydrogel-based 3D culture is an effective strategy for large-scale MSC-Exo production.
- 3D-Exo offers a promising therapeutic approach for cerebral ischemia by reducing neuroinflammation and promoting angiogenesis.
- This study presents a novel exosome-based delivery system for targeting cerebral ischemia.
