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조회192

강우영Wu Young Kang

인천대학교

Biomater. Res., 2024, 28 0005 | https://doi.org/10.34133/bmr.0005 Lyoprotectant Constituents Suited for Lyophilization and Reconstitution of Stem-Cell-Derived Extracellular Vesicles

Wu Young Kang ^1,2, Eun Kyoung Shin ³, Eun Hee Kim³, Min-Ho Kang ^4,5, Chi Young Bang ⁶, Oh Young Bang ^3,7, Jae Min Cha^1,2*

¹Department of Biomedical & Robotics Engineering, College of Engineering, Incheon National University, Incheon 22012, Republic of Korea.

²3D Stem Cell Bioengineering Laboratory, Research Institute for Engineering and Technology, Incheon National University, Incheon 22012, Republic of Korea.

³S&E bio Co., Ltd., Seoul 06351, Republic of Korea.

⁴Department of BioMedical-Chemical Engineering (BMCE), The Catholic University of Korea, Bucheon 14662, Republic of Korea.

⁵Department of Biotechnology, The Catholic University of Korea, Bucheon 14662, Republic of Korea.

⁶Department of Plastic and Reconstructive Surgery, Kangwon National University Hospital, Chuncheon 24341, Republic of Korea.

⁷Department of Neurology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul 06351, Republic of Korea.

*Address correspondence to: Jae Min Cha

Abstract

Stem-cell-derived extracellular vesicles (EVs) are emerging as an alternative approach to stem cell therapy. Successful lyophilization of EVs could enable convenient storage and distribution of EV medicinal products at room temperature for long periods, thus considerably increasing the accessibility of EV therapeutics to patients. In this study, we aimed to identify an appropriate lyoprotectant composition for the lyophilization and reconstitution of stem-cell-derived EVs. MSC-derived EVs were lyophilized using different lyoprotectants, such as dimethyl sulfoxide, mannitol, trehalose, and sucrose, at varying concentrations. Our results revealed that a mixture of trehalose and sucrose at high concentrations could support the formation of amorphous ice by enriching the amorphous phase of the solution, which successfully inhibited the acceleration of buffer component crystallization during lyophilization. Lyophilized and reconstituted EVs were thoroughly evaluated for concentration and size, morphology, and protein and RNA content. The therapeutic effects of the reconstituted EVs were examined using a tube formation assay with human umbilical vein endothelial cells. After rehydration of the lyophilized EVs, most of their generic characteristics were well-maintained, and their therapeutic capacity recovered to levels similar to those of freshly collected EVs. The concentrations and morphologies of the lyophilized EVs were similar to the initial features of the fresh EV group until day 30 at room temperature, although their therapeutic capacity appeared to decrease after 7 days. Our study suggests an appropriate composition of lyoprotectants, particularly for EV lyophilization, which could encourage the applications of stem-cell-derived EV therapeutics in the health industry.

논문정보

형식Research article
게재일2024년 02월 (BRIC 등록일 2024-02-20)
연구진국내 연구진
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