Antioxidant or Apoptosis Inhibitor Supplementation in Culture Media Improves Post-Thaw Recovery of Murine Spermatogonial Stem Cells
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 de...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-05-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/10/5/754 |
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| author | Sang-Eun Jung Hui-Jo Oh Jin-Seop Ahn Yong-Hee Kim Bang-Jin Kim Buom-Yong Ryu |
| author_facet | Sang-Eun Jung Hui-Jo Oh Jin-Seop Ahn Yong-Hee Kim Bang-Jin Kim Buom-Yong Ryu |
| author_sort | Sang-Eun Jung |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-10T11:34:58Z |
| format | Article |
| id | doaj.art-c510300438a149cf859974959302774b |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-10T11:34:58Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-c510300438a149cf859974959302774b2023-11-21T18:57:48ZengMDPI AGAntioxidants2076-39212021-05-0110575410.3390/antiox10050754Antioxidant or Apoptosis Inhibitor Supplementation in Culture Media Improves Post-Thaw Recovery of Murine Spermatogonial Stem CellsSang-Eun Jung0Hui-Jo Oh1Jin-Seop Ahn2Yong-Hee Kim3Bang-Jin Kim4Buom-Yong Ryu5Department of Animal Science and Technology, Chung-Ang University, Anseong-si 17546, Gyeonggi-do, KoreaDepartment of Animal Science and Technology, Chung-Ang University, Anseong-si 17546, Gyeonggi-do, KoreaDepartment of Animal Science and Technology, Chung-Ang University, Anseong-si 17546, Gyeonggi-do, KoreaDepartment of Animal Science and Technology, Chung-Ang University, Anseong-si 17546, Gyeonggi-do, KoreaDepartment of Cancer Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USADepartment of Animal Science and Technology, Chung-Ang University, Anseong-si 17546, Gyeonggi-do, KoreaWe 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.https://www.mdpi.com/2076-3921/10/5/754permatogonial stem cellspost-thaw recoveryreactive oxygen speciesantioxidantapoptosis |
| spellingShingle | Sang-Eun Jung Hui-Jo Oh Jin-Seop Ahn Yong-Hee Kim Bang-Jin Kim Buom-Yong Ryu Antioxidant or Apoptosis Inhibitor Supplementation in Culture Media Improves Post-Thaw Recovery of Murine Spermatogonial Stem Cells Antioxidants permatogonial stem cells post-thaw recovery reactive oxygen species antioxidant apoptosis |
| title | Antioxidant or Apoptosis Inhibitor Supplementation in Culture Media Improves Post-Thaw Recovery of Murine Spermatogonial Stem Cells |
| title_full | Antioxidant or Apoptosis Inhibitor Supplementation in Culture Media Improves Post-Thaw Recovery of Murine Spermatogonial Stem Cells |
| title_fullStr | Antioxidant or Apoptosis Inhibitor Supplementation in Culture Media Improves Post-Thaw Recovery of Murine Spermatogonial Stem Cells |
| title_full_unstemmed | Antioxidant or Apoptosis Inhibitor Supplementation in Culture Media Improves Post-Thaw Recovery of Murine Spermatogonial Stem Cells |
| title_short | Antioxidant or Apoptosis Inhibitor Supplementation in Culture Media Improves Post-Thaw Recovery of Murine Spermatogonial Stem Cells |
| title_sort | antioxidant or apoptosis inhibitor supplementation in culture media improves post thaw recovery of murine spermatogonial stem cells |
| topic | permatogonial stem cells post-thaw recovery reactive oxygen species antioxidant apoptosis |
| url | https://www.mdpi.com/2076-3921/10/5/754 |
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