Comparing the effects of a commercial and a prototype vitrification medium on meiotic spindle morphology and survival rate of mouse oocytes
To compare oocyte survival and meiotic spindle normality between vitrified-warmed oocytes in a mouse embryo assay using Tvitri-4 or Ingámed vitrification media. C57BL/6 female mice aged 8-12 weeks were submitted to superovulation with pregnant mare's serum gonadotropin and human chorionic gonad...
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| Veröffentlicht in: | JBRA assisted reproduction 2022, Vol.26 (3), p.500-507 |
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| creator | Viana, Iara Gonçalves Roberto Vireque, Alessandra Aparecida Navarro, Paula Andrea |
| description | To compare oocyte survival and meiotic spindle normality between vitrified-warmed oocytes in a mouse embryo assay using Tvitri-4 or Ingámed vitrification media.
C57BL/6 female mice aged 8-12 weeks were submitted to superovulation with pregnant mare's serum gonadotropin and human chorionic gonadotropin (hCG) for obtaining of in vivo matured oocytes. The oocytes were randomly distributed into one of the following three groups: CTR - control (fresh oocytes); ING - oocytes vitrified-warmed in a standard commercial kit supplied by Ingámed, and T4 - oocytes vitrified-warmed in the novel prototype Tvitri-4 medium. After warming and recovery culture, oocytes were assessed with respect to survival rate (SR) and both meiotic spindle morphology and chromosome alignment of each oocyte fixed in the sagittal position after immunostaining and analysis by confocal microscopy.
A total of 354 mature oocytes were vitrified in ING (n=178) and T4 (n=176), out of which 299 (85%) survived after warming. Oocyte survival rates were not statistically different (p=0.08) between ING (145/178=81.5%) and T4 (154/176=87.5%). Regarding meiotic normality, there were no significant changes in the proportion of oocytes with normal meiotic spindle morphology and chromosome structure between ING (52,2%) and T4 (63.4%) after warming (RR: 0.95, 95% CI: 0.92-1.607). When the meiotic normality was assessed using the CTR group as a reference in the analysis of relative risk, no significant differences were observed between T4 (63.4%) and CTR (70.5%) (RR: 0.95, 95% CI: 0.72-1.12). On the other hand, the percentage of oocytes retaining normal meiotic spindle morphology and chromosome configuration in ING (52.2%) was lower than in the CTR group (RR: 0.95, 95% CI: 0.57-0.97).
The novel prototype Tvitri-4 medium was efficient in preserve survival rate and meiotic spindle normality of oocytes and, with further verification, may be able to replace commercially available media in future clinical applications. |
| doi_str_mv | 10.5935/1518-0557.20210117 |
| format | Article |
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C57BL/6 female mice aged 8-12 weeks were submitted to superovulation with pregnant mare's serum gonadotropin and human chorionic gonadotropin (hCG) for obtaining of in vivo matured oocytes. The oocytes were randomly distributed into one of the following three groups: CTR - control (fresh oocytes); ING - oocytes vitrified-warmed in a standard commercial kit supplied by Ingámed, and T4 - oocytes vitrified-warmed in the novel prototype Tvitri-4 medium. After warming and recovery culture, oocytes were assessed with respect to survival rate (SR) and both meiotic spindle morphology and chromosome alignment of each oocyte fixed in the sagittal position after immunostaining and analysis by confocal microscopy.
A total of 354 mature oocytes were vitrified in ING (n=178) and T4 (n=176), out of which 299 (85%) survived after warming. Oocyte survival rates were not statistically different (p=0.08) between ING (145/178=81.5%) and T4 (154/176=87.5%). Regarding meiotic normality, there were no significant changes in the proportion of oocytes with normal meiotic spindle morphology and chromosome structure between ING (52,2%) and T4 (63.4%) after warming (RR: 0.95, 95% CI: 0.92-1.607). When the meiotic normality was assessed using the CTR group as a reference in the analysis of relative risk, no significant differences were observed between T4 (63.4%) and CTR (70.5%) (RR: 0.95, 95% CI: 0.72-1.12). On the other hand, the percentage of oocytes retaining normal meiotic spindle morphology and chromosome configuration in ING (52.2%) was lower than in the CTR group (RR: 0.95, 95% CI: 0.57-0.97).
The novel prototype Tvitri-4 medium was efficient in preserve survival rate and meiotic spindle normality of oocytes and, with further verification, may be able to replace commercially available media in future clinical applications.</description><identifier>ISSN: 1518-0557</identifier><identifier>ISSN: 1517-5693</identifier><identifier>EISSN: 1518-0557</identifier><identifier>DOI: 10.5935/1518-0557.20210117</identifier><identifier>PMID: 35261222</identifier><language>eng</language><publisher>Brazil: Sociedade Brasileira de Reprodução Humana (Brazilian Society of Assisted Reproduction)</publisher><subject>Animals ; Cryopreservation ; Cryopreservation - methods ; Female ; Females ; Horses ; Humans ; In vitro fertilization ; In Vitro Oocyte Maturation Techniques - methods ; Mice ; Mice, Inbred C57BL ; Morphology ; Oocytes ; Original ; Pregnancy ; Proteins ; Spindle Apparatus ; Sucrose ; Survival Rate ; Vitrification</subject><ispartof>JBRA assisted reproduction, 2022, Vol.26 (3), p.500-507</ispartof><rights>2022. This work is published under https://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><orcidid>0000-0003-2368-4188</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9355432/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9355432/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,728,781,785,886,4025,27928,27929,27930,53796,53798</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35261222$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Viana, Iara Gonçalves Roberto</creatorcontrib><creatorcontrib>Vireque, Alessandra Aparecida</creatorcontrib><creatorcontrib>Navarro, Paula Andrea</creatorcontrib><title>Comparing the effects of a commercial and a prototype vitrification medium on meiotic spindle morphology and survival rate of mouse oocytes</title><title>JBRA assisted reproduction</title><addtitle>JBRA Assist Reprod</addtitle><description>To compare oocyte survival and meiotic spindle normality between vitrified-warmed oocytes in a mouse embryo assay using Tvitri-4 or Ingámed vitrification media.
C57BL/6 female mice aged 8-12 weeks were submitted to superovulation with pregnant mare's serum gonadotropin and human chorionic gonadotropin (hCG) for obtaining of in vivo matured oocytes. The oocytes were randomly distributed into one of the following three groups: CTR - control (fresh oocytes); ING - oocytes vitrified-warmed in a standard commercial kit supplied by Ingámed, and T4 - oocytes vitrified-warmed in the novel prototype Tvitri-4 medium. After warming and recovery culture, oocytes were assessed with respect to survival rate (SR) and both meiotic spindle morphology and chromosome alignment of each oocyte fixed in the sagittal position after immunostaining and analysis by confocal microscopy.
A total of 354 mature oocytes were vitrified in ING (n=178) and T4 (n=176), out of which 299 (85%) survived after warming. Oocyte survival rates were not statistically different (p=0.08) between ING (145/178=81.5%) and T4 (154/176=87.5%). Regarding meiotic normality, there were no significant changes in the proportion of oocytes with normal meiotic spindle morphology and chromosome structure between ING (52,2%) and T4 (63.4%) after warming (RR: 0.95, 95% CI: 0.92-1.607). When the meiotic normality was assessed using the CTR group as a reference in the analysis of relative risk, no significant differences were observed between T4 (63.4%) and CTR (70.5%) (RR: 0.95, 95% CI: 0.72-1.12). On the other hand, the percentage of oocytes retaining normal meiotic spindle morphology and chromosome configuration in ING (52.2%) was lower than in the CTR group (RR: 0.95, 95% CI: 0.57-0.97).
The novel prototype Tvitri-4 medium was efficient in preserve survival rate and meiotic spindle normality of oocytes and, with further verification, may be able to replace commercially available media in future clinical applications.</description><subject>Animals</subject><subject>Cryopreservation</subject><subject>Cryopreservation - methods</subject><subject>Female</subject><subject>Females</subject><subject>Horses</subject><subject>Humans</subject><subject>In vitro fertilization</subject><subject>In Vitro Oocyte Maturation Techniques - methods</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Morphology</subject><subject>Oocytes</subject><subject>Original</subject><subject>Pregnancy</subject><subject>Proteins</subject><subject>Spindle Apparatus</subject><subject>Sucrose</subject><subject>Survival Rate</subject><subject>Vitrification</subject><issn>1518-0557</issn><issn>1517-5693</issn><issn>1518-0557</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><recordid>eNpdUctqHDEQFCEmNmv_QA5G4Esu6-g5mrkYwhI7AUMuyVloNa1dmdFoLGkW9hv809H6sTg5ddGqLnV1IfSZkmvZcfmVStouiZTqmhFGCaXqAzo7Nj--w6foIucHQgjtKOOCfEKnXLKGMsbO0NMqhskkP25w2QIG58CWjKPDBtsYAiTrzYDN2NfGlGKJZT8B3vmSvPPWFB9HHKD3c8DPyMfiLc6TH_sBcIhp2sYhbvbPEnlOO7-reskUOHwS4pwriHZfIJ-jE2eGDBevdYH-3H7_vfqxvP9193P17X5peUvLEjoLiva9YExAL4VqrGkdE66DjhND16JhorXKyqZdC0cdsabnjZK9ayyXii_QzYvuNK_r6hbGksygp-SDSXsdjdf_vox-qzdxp-vdpeCsCnx5FUjxcYZcdPDZwjCYEaohzRquZMs5l5V69R_1Ic5prPY0U6RVTLFKXCD2wrIp5pzAHZehRB_i1oc09SFN_RZ3Hbp8b-M48hYu_wt5UqiL</recordid><startdate>2022</startdate><enddate>2022</enddate><creator>Viana, Iara Gonçalves Roberto</creator><creator>Vireque, Alessandra Aparecida</creator><creator>Navarro, Paula Andrea</creator><general>Sociedade Brasileira de Reprodução Humana (Brazilian Society of Assisted Reproduction)</general><general>Brazilian Society of Assisted Reproduction</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-2368-4188</orcidid></search><sort><creationdate>2022</creationdate><title>Comparing the effects of a commercial and a prototype vitrification medium on meiotic spindle morphology and survival rate of mouse oocytes</title><author>Viana, Iara Gonçalves Roberto ; Vireque, Alessandra Aparecida ; Navarro, Paula Andrea</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c381t-e9ce71dd4224ed5476ca8f24f9e930a1b46248c7c568b4f1f0cad3675df6c3573</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Animals</topic><topic>Cryopreservation</topic><topic>Cryopreservation - methods</topic><topic>Female</topic><topic>Females</topic><topic>Horses</topic><topic>Humans</topic><topic>In vitro fertilization</topic><topic>In Vitro Oocyte Maturation Techniques - methods</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Morphology</topic><topic>Oocytes</topic><topic>Original</topic><topic>Pregnancy</topic><topic>Proteins</topic><topic>Spindle Apparatus</topic><topic>Sucrose</topic><topic>Survival Rate</topic><topic>Vitrification</topic><toplevel>online_resources</toplevel><creatorcontrib>Viana, Iara Gonçalves Roberto</creatorcontrib><creatorcontrib>Vireque, Alessandra Aparecida</creatorcontrib><creatorcontrib>Navarro, Paula Andrea</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>JBRA assisted reproduction</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Viana, Iara Gonçalves Roberto</au><au>Vireque, Alessandra Aparecida</au><au>Navarro, Paula Andrea</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparing the effects of a commercial and a prototype vitrification medium on meiotic spindle morphology and survival rate of mouse oocytes</atitle><jtitle>JBRA assisted reproduction</jtitle><addtitle>JBRA Assist Reprod</addtitle><date>2022</date><risdate>2022</risdate><volume>26</volume><issue>3</issue><spage>500</spage><epage>507</epage><pages>500-507</pages><issn>1518-0557</issn><issn>1517-5693</issn><eissn>1518-0557</eissn><abstract>To compare oocyte survival and meiotic spindle normality between vitrified-warmed oocytes in a mouse embryo assay using Tvitri-4 or Ingámed vitrification media.
C57BL/6 female mice aged 8-12 weeks were submitted to superovulation with pregnant mare's serum gonadotropin and human chorionic gonadotropin (hCG) for obtaining of in vivo matured oocytes. The oocytes were randomly distributed into one of the following three groups: CTR - control (fresh oocytes); ING - oocytes vitrified-warmed in a standard commercial kit supplied by Ingámed, and T4 - oocytes vitrified-warmed in the novel prototype Tvitri-4 medium. After warming and recovery culture, oocytes were assessed with respect to survival rate (SR) and both meiotic spindle morphology and chromosome alignment of each oocyte fixed in the sagittal position after immunostaining and analysis by confocal microscopy.
A total of 354 mature oocytes were vitrified in ING (n=178) and T4 (n=176), out of which 299 (85%) survived after warming. Oocyte survival rates were not statistically different (p=0.08) between ING (145/178=81.5%) and T4 (154/176=87.5%). Regarding meiotic normality, there were no significant changes in the proportion of oocytes with normal meiotic spindle morphology and chromosome structure between ING (52,2%) and T4 (63.4%) after warming (RR: 0.95, 95% CI: 0.92-1.607). When the meiotic normality was assessed using the CTR group as a reference in the analysis of relative risk, no significant differences were observed between T4 (63.4%) and CTR (70.5%) (RR: 0.95, 95% CI: 0.72-1.12). On the other hand, the percentage of oocytes retaining normal meiotic spindle morphology and chromosome configuration in ING (52.2%) was lower than in the CTR group (RR: 0.95, 95% CI: 0.57-0.97).
The novel prototype Tvitri-4 medium was efficient in preserve survival rate and meiotic spindle normality of oocytes and, with further verification, may be able to replace commercially available media in future clinical applications.</abstract><cop>Brazil</cop><pub>Sociedade Brasileira de Reprodução Humana (Brazilian Society of Assisted Reproduction)</pub><pmid>35261222</pmid><doi>10.5935/1518-0557.20210117</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0003-2368-4188</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | JBRA assisted reproduction, 2022, Vol.26 (3), p.500-507 |
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| subjects | Animals Cryopreservation Cryopreservation - methods Female Females Horses Humans In vitro fertilization In Vitro Oocyte Maturation Techniques - methods Mice Mice, Inbred C57BL Morphology Oocytes Original Pregnancy Proteins Spindle Apparatus Sucrose Survival Rate Vitrification |
| title | Comparing the effects of a commercial and a prototype vitrification medium on meiotic spindle morphology and survival rate of mouse oocytes |
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