Hdl Handle:
http://hdl.handle.net/10675.2/684
Title:
Comparison and Avoidance of Toxicity of Penetrating Cryoprotectants
Authors:
Szurek, Edyta A.; Eroglu, Ali
Abstract:
The objective of this study was to elucidate the toxicity of widely used penetrating cryoprotective agents (CPAs) to mammalian oocytes. To this end, mouse metaphase II (M II) oocytes were exposed to 1.5 M solutions of dimethylsulfoxide (DMSO), ethylene glycol (EG), or propanediol (PROH) prepared in phosphate buffered saline (PBS) containing 10% fetal bovine serum. To address the time- and temperature-dependence of the CPA toxicity, M II oocytes were exposed to the aforementioned CPAs at room temperature (RT, ,23uC) and 37uC for 15 or 30 minutes. Subsequently, the toxicity of each CPA was evaluated by examining post-exposure survival, fertilization, embryonic development, chromosomal abnormalities, and parthenogenetic activation of treated oocytes. Untreated oocytes served as controls. Exposure of MII oocytes to 1.5 M DMSO or 1.5 M EG at RT for 15 min did not adversely affect any of the evaluated criteria. In contrast, 1.5 M PROH induced a significant increase in oocyte degeneration (54.2%) and parthenogenetic activation (16%) under same conditions. When the CPA exposure was performed at 37uC, the toxic effect of PROH further increased, resulting in lower survival (15%) and no fertilization while the toxicity of DMSO and EG was still insignificant. Nevertheless, it was possible to completely avoid the toxicity of PROH by decreasing its concentration to 0.75 M and combining it with 0.75 M DMSO to bring the total CPA concentration to a cryoprotective level. Moreover, combining lower concentrations (i.e., 0.75 M) of PROH and DMSO significantly improved the cryosurvival of MII oocytes compared to the equivalent concentration of DMSO alone. Taken together, our results suggest that from the perspective of CPA toxicity, DMSO and EG are safer to use in slow cooling protocols while a lower concentration of PROH can be combined with another CPA to avoid its toxicity and to improve the cryosurvival as well.
Citation:
PLoS One. 2011 Nov 16; 6(11):e27604
Issue Date:
16-Nov-2011
URI:
http://hdl.handle.net/10675.2/684
DOI:
10.1371/journal.pone.0027604
PubMed ID:
22110685
PubMed Central ID:
PMC3217997
Type:
Article
ISSN:
1932-6203
Appears in Collections:
Institute of Molecular Medicine and Genetics: Faculty Research and Presentations

Full metadata record

DC FieldValue Language
dc.contributor.authorSzurek, Edyta A.en_US
dc.contributor.authorEroglu, Alien_US
dc.date.accessioned2012-10-26T16:29:32Z-
dc.date.available2012-10-26T16:29:32Z-
dc.date.issued2011-11-16en_US
dc.identifier.citationPLoS One. 2011 Nov 16; 6(11):e27604en_US
dc.identifier.issn1932-6203en_US
dc.identifier.pmid22110685en_US
dc.identifier.doi10.1371/journal.pone.0027604en_US
dc.identifier.urihttp://hdl.handle.net/10675.2/684-
dc.description.abstractThe objective of this study was to elucidate the toxicity of widely used penetrating cryoprotective agents (CPAs) to mammalian oocytes. To this end, mouse metaphase II (M II) oocytes were exposed to 1.5 M solutions of dimethylsulfoxide (DMSO), ethylene glycol (EG), or propanediol (PROH) prepared in phosphate buffered saline (PBS) containing 10% fetal bovine serum. To address the time- and temperature-dependence of the CPA toxicity, M II oocytes were exposed to the aforementioned CPAs at room temperature (RT, ,23uC) and 37uC for 15 or 30 minutes. Subsequently, the toxicity of each CPA was evaluated by examining post-exposure survival, fertilization, embryonic development, chromosomal abnormalities, and parthenogenetic activation of treated oocytes. Untreated oocytes served as controls. Exposure of MII oocytes to 1.5 M DMSO or 1.5 M EG at RT for 15 min did not adversely affect any of the evaluated criteria. In contrast, 1.5 M PROH induced a significant increase in oocyte degeneration (54.2%) and parthenogenetic activation (16%) under same conditions. When the CPA exposure was performed at 37uC, the toxic effect of PROH further increased, resulting in lower survival (15%) and no fertilization while the toxicity of DMSO and EG was still insignificant. Nevertheless, it was possible to completely avoid the toxicity of PROH by decreasing its concentration to 0.75 M and combining it with 0.75 M DMSO to bring the total CPA concentration to a cryoprotective level. Moreover, combining lower concentrations (i.e., 0.75 M) of PROH and DMSO significantly improved the cryosurvival of MII oocytes compared to the equivalent concentration of DMSO alone. Taken together, our results suggest that from the perspective of CPA toxicity, DMSO and EG are safer to use in slow cooling protocols while a lower concentration of PROH can be combined with another CPA to avoid its toxicity and to improve the cryosurvival as well.en_US
dc.rightsSzurek, Eroglu. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_US
dc.subjectResearch Articleen_US
dc.subjectBiologyen_US
dc.subjectAnatomy and Physiologyen_US
dc.subjectReproductive Systemen_US
dc.subjectSexual Reproductionen_US
dc.subjectCryobiologyen_US
dc.subjectDevelopmental Biologyen_US
dc.subjectEmbryologyen_US
dc.subjectModel Organismsen_US
dc.subjectAnimal Modelsen_US
dc.subjectMouseen_US
dc.subjectMedicineen_US
dc.subjectAnatomy and Physiologyen_US
dc.subjectReproductive Systemen_US
dc.subjectReproductive Physiologyen_US
dc.subjectObstetrics and Gynecologyen_US
dc.subjectFemale Subfertilityen_US
dc.subjectWomen's Healthen_US
dc.subject.meshAnimalsen_US
dc.subject.meshCell Survivalen_US
dc.subject.meshChromosome Aberrationsen_US
dc.subject.meshCryoprotective Agentsen_US
dc.subject.meshDose-Response Relationship, Drugen_US
dc.subject.meshEmbryonic Developmenten_US
dc.subject.meshFertilizationen_US
dc.subject.meshMiceen_US
dc.subject.meshOocytesen_US
dc.subject.meshPloidiesen_US
dc.subject.meshTemperatureen_US
dc.subject.meshTime Factorsen_US
dc.subject.meshToxicity Testsen_US
dc.titleComparison and Avoidance of Toxicity of Penetrating Cryoprotectantsen_US
dc.typeArticleen_US
dc.identifier.pmcidPMC3217997en_US
dc.contributor.corporatenameInstitute of Molecular Medicine and Genetics-
dc.contributor.corporatenameDepartment of Medicine-
dc.contributor.corporatenameDepartment of Obstetrics and Gynecology-
dc.contributor.corporatenameGHSU Cancer Center-
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