한빛사 논문
Kyung Min Lim a,f,1, Sehee Kim a,1, Jeonghun Yeom b,1, Yujin Choi a, Yoonjoo Lee a, Jongyub An a, Minchan Gil a, Ahmed Abdal Dayem a, Kyeongseok Kim a, Geun-Ho Kang a,f, Aram Kim c, Kwonho Hong a, Kyunggon Kim b,d,e,⇑, Ssang-Goo Cho a,f,⇑
aDepartment of Stem Cell & Regenerative Biotechnology and Institute of Advanced Regenerative Science, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea
bConvergence Medicine Research Center, Asan Institute for Life Sciences, Asan Medical Center, 88-gil, 43 Olympic-ro, Songpa-gu, Seoul 05505, Republic of Korea
cDepartment of Urology, Konkuk University Medical Center, Konkuk University School of Medicine, Seoul 05029, Republic of Korea
dBiomedical Research Center, Asan Institute for Life Sciences, Asan Medical Center, 88-gil, 43 Olympic-ro, Songpa-gu, Seoul 05505, Republic of Korea
eDepartment of Convergence Medicine, University of Ulsan College of Medicine, 88-gil, 43 Olympic-ro, Songpa-gu, Seoul 05505, Republic of Korea
fR&D Team, StemExOne Co., Ltd., 303, Life Science Bldg, 120, Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea
⇑ Corresponding authors
1 These authors contributed equally.
Abstract
Introduction
Mesenchymal stromal cells (MSCs) release extracellular vesicles (MSC-EVs) containing various cargoes. Although MSC-EVs show significant therapeutic effects, the low production of EVs in MSCs hinders MSC-EV-mediated therapeutic development.
Objectives
Here, we developed an advanced three-dimensional (a3D) dynamic culture technique with exogenous transforming growth factor beta-3 (TGF-β3) treatment (T-a3D) to produce potent MSC-EVs.
Methods
Our system enabled preparation of a highly concentrated EV-containing medium for efficient EV isolation and purification with higher yield and efficacy.
Results
MSC spheroids in T-a3D system (T-a3D spheroids) showed high expression of CD9 and TGF-β3, which was dependent on TGF-β signaling. Treatment with EVs produced under T-a3D conditions (T-a3D-EVs) led to significantly improved migration of dermal fibroblasts and wound closure in an excisional wound model. The relative total efficacy (relative yield of single-batch EVs (10–11-fold) × relative regeneration effect of EVs (2–3-fold)) of T-a3D-EVs was approximately up to 33-fold higher than that of 2D-EVs. Importantly the quantitative proteomic analyses of the T-a3D spheroids and T-a3D-EVs supported the improved EV production as well as the therapeutic potency of T-a3D-EVs.
Conclusion
TGF-β signalling differentially regulated by fluid shear stress produced in our system and exogenous TGF-β3 addition was confirmed to play an important role in the enhanced production of EVs with modified protein cargoes. We suggest that the T-a3D system leads to the efficient production of MSC-EVs with high potential in therapies and clinical development.
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