Cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells.
Development of techniques to isolate, culture, and transplant human spermatogonial stem cells (SSCs) has the future potential to treat male infertility. To maximize the efficiency of these techniques, methods for SSC cryopreservation need to be developed to bank SSCs for extended periods of time. Al...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3551902?pdf=render |
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| author | Yong-An Lee Yong-Hee Kim Bang-Jin Kim Byung-Gak Kim Ki-Jung Kim Joong-Hyuck Auh Jonathan A Schmidt Buom-Yong Ryu |
| author_facet | Yong-An Lee Yong-Hee Kim Bang-Jin Kim Byung-Gak Kim Ki-Jung Kim Joong-Hyuck Auh Jonathan A Schmidt Buom-Yong Ryu |
| author_sort | Yong-An Lee |
| collection | DOAJ |
| description | Development of techniques to isolate, culture, and transplant human spermatogonial stem cells (SSCs) has the future potential to treat male infertility. To maximize the efficiency of these techniques, methods for SSC cryopreservation need to be developed to bank SSCs for extended periods of time. Although, it has been demonstrated that SSCs can reinitiate spermatogenesis after freezing, optimal cryopreservation protocols that maximize SSC proliferative capacity post-thaw have not been identified. The objective of this study was to develop an efficient cryopreservation technique for preservation of SSCs. To identify efficient cryopreservation methods for long-term preservation of SSCs, isolated testis cells enriched for SSCs were placed in medium containing dimethyl sulfoxide (DMSO) or DMSO and trehalose (50 mM, 100 mM, or 200 mM), and frozen in liquid nitrogen for 1 week, 1 month, or 3 months. Freezing in 50 mM trehalose resulted in significantly higher cell viability compared to DMSO at all thawing times and a higher proliferation rate compared to DMSO for the 1 week freezing period. Freezing in 200 mM trehalose did not result in increased cell viability; however, proliferation activity was significantly higher and percentage of apoptotic cells was significantly lower compared to DMSO after freezing for 1 and 3 months. To confirm the functionality of SSCs frozen in 200 mM trehalose, SSC transplantation was performed. Donor SSCs formed spermatogenic colonies and sperm capable of generating normal progeny. Collectively, these results indicate that freezing in DMSO with 200 mM trehalose serves as an efficient method for the cryopreservation of SSCs. |
| first_indexed | 2024-12-14T01:02:16Z |
| format | Article |
| id | doaj.art-821a83bb63da46a18875970c0acca8cb |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T01:02:16Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-821a83bb63da46a18875970c0acca8cb2022-12-21T23:23:11ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0181e5488910.1371/journal.pone.0054889Cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells.Yong-An LeeYong-Hee KimBang-Jin KimByung-Gak KimKi-Jung KimJoong-Hyuck AuhJonathan A SchmidtBuom-Yong RyuDevelopment of techniques to isolate, culture, and transplant human spermatogonial stem cells (SSCs) has the future potential to treat male infertility. To maximize the efficiency of these techniques, methods for SSC cryopreservation need to be developed to bank SSCs for extended periods of time. Although, it has been demonstrated that SSCs can reinitiate spermatogenesis after freezing, optimal cryopreservation protocols that maximize SSC proliferative capacity post-thaw have not been identified. The objective of this study was to develop an efficient cryopreservation technique for preservation of SSCs. To identify efficient cryopreservation methods for long-term preservation of SSCs, isolated testis cells enriched for SSCs were placed in medium containing dimethyl sulfoxide (DMSO) or DMSO and trehalose (50 mM, 100 mM, or 200 mM), and frozen in liquid nitrogen for 1 week, 1 month, or 3 months. Freezing in 50 mM trehalose resulted in significantly higher cell viability compared to DMSO at all thawing times and a higher proliferation rate compared to DMSO for the 1 week freezing period. Freezing in 200 mM trehalose did not result in increased cell viability; however, proliferation activity was significantly higher and percentage of apoptotic cells was significantly lower compared to DMSO after freezing for 1 and 3 months. To confirm the functionality of SSCs frozen in 200 mM trehalose, SSC transplantation was performed. Donor SSCs formed spermatogenic colonies and sperm capable of generating normal progeny. Collectively, these results indicate that freezing in DMSO with 200 mM trehalose serves as an efficient method for the cryopreservation of SSCs.http://europepmc.org/articles/PMC3551902?pdf=render |
| spellingShingle | Yong-An Lee Yong-Hee Kim Bang-Jin Kim Byung-Gak Kim Ki-Jung Kim Joong-Hyuck Auh Jonathan A Schmidt Buom-Yong Ryu Cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells. PLoS ONE |
| title | Cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells. |
| title_full | Cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells. |
| title_fullStr | Cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells. |
| title_full_unstemmed | Cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells. |
| title_short | Cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells. |
| title_sort | cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells |
| url | http://europepmc.org/articles/PMC3551902?pdf=render |
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