Abstract:
RESEARCH QUESTION:From a clinical perspective, which parameters grant optimal cryopreservation of mouse testicular cell suspensions? DESIGN:We studied the effect of different cryopreservation rates, the addition of sugars, different vessels and the addition of an apoptotic inhibitor on the efficiency of testicular cell suspension cryopreservation. After thawing and warming, testicular cell suspensions were transplanted to recipient mice for further functional assay. After selecting the optimal cryopreservation procedure, a second experiment compared the transplantation efficiency between the selected freezing protocol and fresh testicular cell suspensions. RESULTS:Multiple- and single-step freezing did not differ significantly in terms of recovered viable cells (RVC) (33 ± 28% and 38 ± 25%). The addition of sucrose did not result in a higher RVC (33 ± 20%). Cells frozen in vials recovered better than those frozen in straws (52 ± 20% versus 33 ± 20%; P = 0.0049). The inclusion of an apoptosis inhibitor (z-VAD[Oe]-FMK) significantly increased the RVC after thawing (61 ± 18% versus 50 ± 17%; P = 0.0480). When comparing the optimal cryopreservation procedure with fresh testicular cell suspensions, a lower RVC (63 ± 11% versus 92 ± 4%; P < 0.0001) and number of donor-derived spermatogonial stem cell colonies per testis (34.04 ± 2.34 versus 16.78 ± 7.76; P = 0.0051) were observed. CONCLUSION:Upon freeze-thawing or vitrification-warming, and assessment of donor-derived spermatogenesis after transplantation, Dulbecco's modified Eagle's medium supplemented with 1.5M dimethyl-sulphoxide, 10% fetal calf serum and 60 µM of Z-VAD-(OMe)-FMK in vials at a freezing rate of -1°C/min was optimal.
journal_name
Reprod Biomed Onlinejournal_title
Reproductive biomedicine onlineauthors
Onofre J,Faes K,Kadam P,Vicini E,van Pelt AMM,Goossens Edoi
10.1016/j.rbmo.2018.04.045subject
Has Abstractpub_date
2018-07-01 00:00:00pages
6-17issue
1eissn
1472-6483issn
1472-6491pii
S1472-6483(18)30222-0journal_volume
37pub_type
杂志文章abstract::Artificial oocyte activation using Ca(2+)ionophores or similar compounds is a widely applied technique in IVF laboratories. This is all the more interesting as most of the agents aiming for intracellular Ca(2+) increase do not result in physiological Ca(2+) oscillations but much rather cause a single Ca(2+) transient....
journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
pub_type: 临床试验,杂志文章,随机对照试验
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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doi:10.1016/s1472-6483(10)60146-0
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journal_title:Reproductive biomedicine online
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doi:10.1016/j.rbmo.2018.11.020
更新日期:2019-04-01 00:00:00
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journal_title:Reproductive biomedicine online
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pub_type: 杂志文章,多中心研究,随机对照试验
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更新日期:2019-06-01 00:00:00
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doi:10.1016/j.rbmo.2010.06.033
更新日期:2010-10-01 00:00:00
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journal_title:Reproductive biomedicine online
pub_type: 指南,杂志文章
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更新日期:2008-01-01 00:00:00
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journal_title:Reproductive biomedicine online
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doi:10.1016/s1472-6483(10)61284-9
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journal_title:Reproductive biomedicine online
pub_type: 杂志文章
doi:10.1016/s1472-6483(10)62183-9
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
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journal_title:Reproductive biomedicine online
pub_type: 杂志文章
doi:10.1016/s1472-6483(10)61801-9
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journal_title:Reproductive biomedicine online
pub_type: 杂志文章
doi:10.1016/j.rbmo.2011.03.019
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journal_title:Reproductive biomedicine online
pub_type: 杂志文章
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更新日期:2018-05-01 00:00:00
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journal_title:Reproductive biomedicine online
pub_type: 杂志文章
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更新日期:2011-12-01 00:00:00
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journal_title:Reproductive biomedicine online
pub_type: 评论,杂志文章
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更新日期:2011-12-01 00:00:00
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journal_title:Reproductive biomedicine online
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更新日期:2020-03-01 00:00:00
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