Tuning Hydrogel Mechanics and Microstructure to Maximize Extracellular Vesicle Production from Mesenchymal Stem Cells.

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

• Level A · Stronger Clinical Evidence
• Level B · Emerging clinical evidence with positive signals
• Level C · Early human research exploring benefits
• Level D · Scientific groundwork from lab and animal studies
• Emerging · Emerging topic under active research

Abstract (original English)

Background The secretory output from mesenchymal stem cells (MSCs) have emerged as promising therapeutics with extracellular vesicles (EVs) gaining prominence due to solution stability and optimal size for overcoming biological barriers during delivery. However, reproducible and scalable production of EVs for therapeutic use remains a challenge in biotechnology. Here we demonstrate optimization of EV production from MSCs using soft hydrogel microcarriers. Methods Gelatin methacryloyl (GelMA) hydrogels were prepared at a range of concentrations for the culture of two sources of MSCs: adipose derived stem cells (ADSCs) and induced pluripotent stem cell derived MSCs (iMSCs). The mechanical properties of the hydrogels were evaluated using shear rheology. EVs were isolated and analyzed for physical and biological characteristics using electron microscopy, nanoparticle tracking, proteomics, and functional assays for wound healing and angiogenesis. Results Both cell types were responsive to hydrogel stiffness (0.3-16 KPa), showing optimal EV secretion from cultures on 10 KPa hydrogels, with a further 18-fold increase when formulated as microcarriers compared to traditional monolayer culture. Proteomics analysis and functional assays revealed that EVs from microcarrier culture displayed increased wound healing and regenerative properties. Conclusion This study demonstrates the advantag

Evidence level

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

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