Antioxidant or apoptosis inhibitor supplementation in culture media improves post-thaw recovery of murine spermatogonial stem cellsopen access
- Authors
- Jung, Sang-Eun; Oh, Hui-Jo; Ahn, Jin-Seop; Kim, Yong-Hee; Kim, Bang-Jin; Ryu, Buom-Yong
- Issue Date
- May-2021
- Publisher
- MDPI AG
- Keywords
- Antioxidant; Apoptosis; Permatogonial stem cells; Post-thaw recovery; Reactive oxygen species
- Citation
- Antioxidants, v.10, no.5
- Journal Title
- Antioxidants
- Volume
- 10
- Number
- 5
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/52280
- DOI
- 10.3390/antiox10050754
- ISSN
- 2076-3921
2076-3921
- Abstract
- We postulated that supplementation of antioxidant or apoptosis inhibitor in post-thaw culture media of spermatogonial stem cells (SSCs) alleviates reactive oxygen species (ROS) generation and apoptosis. Our aim was to develop an effective culture media for improving post-thaw recovery of SSCs. To determine the efficacy of supplementation with hypotaurine (HTU), α-tocopherol (α-TCP), and Z-DEVD-FMK (ZDF), we assessed the relative proliferation rate and SSC functional activity and performed a ROS generation assay, apoptosis assay, and western blotting for determination of the Bax/Bcl-xL ratio, as well as immunocytochemistry and real-time quantitative polymerase chain reaction (RT-qPCR) for SSC characterization. The relative proliferation rates with HTU 400 µM (133.7 ± 3.2%), α-TCP 400 µM (158.9 ± 3.6%), and ZDF 200 µM (133.1 ± 7.6%) supplementation were higher than that in the DMSO control (100 ± 3.6%). ROS generation was reduced with α-TCP 400 µM (0.8-fold) supplementation in comparison with the control (1.0-fold). Early apoptosis and Bax/Bcl-xL were lower with α-TCP 400 µM (2.4 ± 0.4% and 0.5-fold) and ZDF 200 µM (1.8 ± 0.4% and 0.3-fold) supplementation in comparison with the control (5.3 ± 1.4% and 1.0-fold) with normal characterization and functional activity. Supplementation of post-thaw culture media with α-TCP 400 µM and ZDF 200 µM improved post-thaw recovery of frozen SSCs via protection from ROS generation and apoptosis after cryo-thawing. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
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