Background: Mesenchymal stem cell-derived conditioned medium (MSC-CM) has emerged as a promising cell-free tool for restoring degenerative diseases and treating traumatic injuries. The present study describes the effect of selenium as a reactive oxygen species (ROS) scavenger and its additive effect with basic fibroblast growth factor (bFGF) on in vitro expansion of amniotic fluid (AF)-MSCs and the paracrine actions of AF-MSC-CM as well as the associated cellular and molecular mechanisms. Methods: In this study, we obtained CM from human AF-MSCs cultured with selenium. The stemness of selenium-treated AF-MSCs was evaluated by cell growth and differentiation potential. Human fibroblasts were treated with AF-MSC-CM and analyzed for cell signaling changes. For in vivo wound healing assay, ICR mice with a full-thickness skin wound were used. Results: Selenium played a critical role in in vitro expansion of AF-MSCs through activation of the AKT-ERK1/2, Smad2, and Stat3 signaling pathways along with inactivation of GSK3β. When administered together with bFGF, it showed remarkable effect in inhibiting ROS accumulation and preserving their multipotency. Proliferation and migration of human dermal fibroblasts and in vivo wound healing were improved in the CMs derived from AF-MSCs exposed to selenium and bFGF, which was caused by the Smad2, AKT-MEK1/2-ERK, and NFκB signaling triggered by the paracrine factors of AF-MSCs, such as TGF-β, VEGF, and IL-6. Our results suggest the following: (a) supplementation of selenium in AF-MSC culture contributes to in vitro expansion and preservation of multipotency, (b) ROS accumulation causes progressive losses in proliferative and differentiation potential, (c) the separate activities of bFGF and selenium in MSCs exert an additive effect when used together, and (d) the additive combination improves the therapeutic effects of AF-MSC-derived CMs on tissue repair and regeneration. Conclusion: Antioxidants, such as selenium, should be considered as an essential supplement for eliciting the paracrine effects of MSC-CMs.
Bibliographical noteFunding Information:
The authors acknowledge the Korea Small and Medium Business Administration, Ministry of Education, Science and Technology, the Ministry of Trade, Industry and Energy (MOTIE), and the Korea Institute for the Advancement of Technology and School of Life Sciences and Biotechnology for BK21 PLUS, Korea University.
This work was supported by grant nos. C0217167 and C0350704 from Business for Cooperative R&D between Industry, Academy, and Research Institute, as funded by the Korea Small and Medium Business Administration in 2012 and 2016 and the Ministry of Trade, Industry and Energy(MOTIE) and the Korea Institute for the Advancement of Technology (KIAT) (N0002405,2017). This work was funded by Stem lab (No. Q1410061), ICT & Future Planning (MSIP), as supported by the Korean government (Ministry of Education, Science and Technology) (NRF-2010-0020347), and School of Life Sciences and Biotechnology for BK21 PLUS, Korea University.
© 2018 The Author(s).
- Amniotic fluid-derived mesenchymal stem cell
- Conditioned media
- Paracrine factors
- Wound healing
ASJC Scopus subject areas
- Medicine (miscellaneous)
- Molecular Medicine
- Biochemistry, Genetics and Molecular Biology (miscellaneous)
- Cell Biology