Effects of three different types of antifreeze proteins on mouse ovarian tissue cryopreservation and transplantation
Ovarian tissue (OT) cryopreservation is effective in preserving fertility in cancer patients who have concerns about fertility loss due to cancer treatment. However, the damage incurred at different steps during the cryopreservation procedure may cause follicular depletion; hence, preventing chillin...
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| Veröffentlicht in: | PloS one 2015-05, Vol.10 (5), p.e0126252-e0126252 |
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| creator | Lee, Jaewang Kim, Seul Ki Youm, Hye Won Kim, Hak Jun Lee, Jung Ryeol Suh, Chang Suk Kim, Seok Hyun |
| description | Ovarian tissue (OT) cryopreservation is effective in preserving fertility in cancer patients who have concerns about fertility loss due to cancer treatment. However, the damage incurred at different steps during the cryopreservation procedure may cause follicular depletion; hence, preventing chilling injury would help maintain ovarian function.
This study was designed to investigate the beneficial effects of different antifreeze proteins (AFPs) on mouse ovarian tissue cryopreservation and transplantation.
Ovaries were obtained from 5-week-old B6D2F1 mice, and each ovary was cryopreserved using two-step vitrification and four-step warming procedures. In Experiment I, ovaries were randomly allocated into fresh, vitrification control, and nine experimental groups according to the AFP type (FfIBP, LeIBP, type III) and concentration (0.1, 1, 10 mg/mL) used. After vitrification and warming, 5,790 ovarian follicles were evaluated using histology and TUNEL assays, and immunofluorescence for τH2AX and Rad51 was used to detect DNA double-strand breaks (DSBs) and repair (DDR), respectively. In Experiment II, 20 mice were randomly divided into two groups: one where the vitrification and warming media were supplemented with 10 mg/mL LeIBP, and the other where media alone were used (control). Ovaries were then autotransplanted under both kidney capsules 7 days after vitrification together with the addition of 10 mg/mL LeIBP in the vitrification-warming media. After transplantation, the ovarian follicles, the percentage of apoptotic follicles, the extent of the CD31-positive area, and the serum FSH levels of the transplanted groups were compared.
In Experiment I, the percentage of total grade 1 follicles was significantly higher in the 10 mg/mL LeIBP group than in the vitrification control, while all AFP-treated groups had significantly improved grade 1 primordial follicle numbers compared with those of the vitrification control. The number of apoptotic (TUNEL-positive) follicles was significantly decreased in the groups treated with 1 and 10 mg/mL LeIBP. The proportion of τH2AX-positive follicles was significantly reduced in all AFP-treated groups, while the proportion of Rad51-positive follicles was significantly decreased in only the FfIBP- and LeIBP-treated groups. In Experiment II, after autotransplantation of OT vitrified with 10 mg/mL of LeIBP, the percentage of total grade 1 and primordial grade 1 follicles, and the extent of the CD31-positive area, were increase |
| doi_str_mv | 10.1371/journal.pone.0126252 |
| format | Article |
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This study was designed to investigate the beneficial effects of different antifreeze proteins (AFPs) on mouse ovarian tissue cryopreservation and transplantation.
Ovaries were obtained from 5-week-old B6D2F1 mice, and each ovary was cryopreserved using two-step vitrification and four-step warming procedures. In Experiment I, ovaries were randomly allocated into fresh, vitrification control, and nine experimental groups according to the AFP type (FfIBP, LeIBP, type III) and concentration (0.1, 1, 10 mg/mL) used. After vitrification and warming, 5,790 ovarian follicles were evaluated using histology and TUNEL assays, and immunofluorescence for τH2AX and Rad51 was used to detect DNA double-strand breaks (DSBs) and repair (DDR), respectively. In Experiment II, 20 mice were randomly divided into two groups: one where the vitrification and warming media were supplemented with 10 mg/mL LeIBP, and the other where media alone were used (control). Ovaries were then autotransplanted under both kidney capsules 7 days after vitrification together with the addition of 10 mg/mL LeIBP in the vitrification-warming media. After transplantation, the ovarian follicles, the percentage of apoptotic follicles, the extent of the CD31-positive area, and the serum FSH levels of the transplanted groups were compared.
In Experiment I, the percentage of total grade 1 follicles was significantly higher in the 10 mg/mL LeIBP group than in the vitrification control, while all AFP-treated groups had significantly improved grade 1 primordial follicle numbers compared with those of the vitrification control. The number of apoptotic (TUNEL-positive) follicles was significantly decreased in the groups treated with 1 and 10 mg/mL LeIBP. The proportion of τH2AX-positive follicles was significantly reduced in all AFP-treated groups, while the proportion of Rad51-positive follicles was significantly decreased in only the FfIBP- and LeIBP-treated groups. In Experiment II, after autotransplantation of OT vitrified with 10 mg/mL of LeIBP, the percentage of total grade 1 and primordial grade 1 follicles, and the extent of the CD31-positive area, were increased significantly. Moreover, the levels of serum FSH and the percentage of TUNEL-positive follicles were significantly lower in the LeIBP-treated than in the control group.
A supplementation with high concentrations of AFPs had protective effects on follicle preservation during OT vitrification-warming procedures. The group treated with LeIBP was protected most effectively. The beneficial effects of LeIBP were also observed after autotransplantation of vitrified-warmed OT. Further studies are necessary to determine the exact mechanism of these protective effects.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0126252</identifier><identifier>PMID: 25938445</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Animals ; Antifreeze proteins ; Antifreeze Proteins - metabolism ; Antifreeze Proteins - pharmacology ; Apoptosis ; Apoptosis - drug effects ; Cancer ; Cancer therapies ; Care and treatment ; Complications and side effects ; Cryopreservation ; Cryopreservation - methods ; Cryoprotective Agents - metabolism ; Cryoprotective Agents - pharmacology ; Deoxyribonucleic acid ; DNA ; DNA damage ; Female ; Fertility ; Follicle Stimulating Hormone - blood ; Follicle-stimulating hormone ; Follicles ; Gynecology ; Health aspects ; Histology ; Immunofluorescence ; Immunohistochemistry ; Injury prevention ; Kidneys ; Leucosporidium ; Medicine ; Mice ; Obstetrics ; Ovarian cancer ; Ovarian Follicle - cytology ; Ovarian Follicle - drug effects ; Ovaries ; Ovary - cytology ; Ovary - transplantation ; Polypeptides ; Proteins ; Quality ; Reproductive status ; Supplements ; Transplantation ; Transplantation, Autologous ; Vascular endothelial growth factor ; Vitrification</subject><ispartof>PloS one, 2015-05, Vol.10 (5), p.e0126252-e0126252</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>2015 Lee et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2015 Lee et al 2015 Lee et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-74b6a69e4f160753e5716de375f22702c42b93c863c1b962f74ed2e6fb4c0783</citedby><cites>FETCH-LOGICAL-c692t-74b6a69e4f160753e5716de375f22702c42b93c863c1b962f74ed2e6fb4c0783</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4418816/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4418816/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25938445$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Schlatt, Stefan</contributor><creatorcontrib>Lee, Jaewang</creatorcontrib><creatorcontrib>Kim, Seul Ki</creatorcontrib><creatorcontrib>Youm, Hye Won</creatorcontrib><creatorcontrib>Kim, Hak Jun</creatorcontrib><creatorcontrib>Lee, Jung Ryeol</creatorcontrib><creatorcontrib>Suh, Chang Suk</creatorcontrib><creatorcontrib>Kim, Seok Hyun</creatorcontrib><title>Effects of three different types of antifreeze proteins on mouse ovarian tissue cryopreservation and transplantation</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Ovarian tissue (OT) cryopreservation is effective in preserving fertility in cancer patients who have concerns about fertility loss due to cancer treatment. However, the damage incurred at different steps during the cryopreservation procedure may cause follicular depletion; hence, preventing chilling injury would help maintain ovarian function.
This study was designed to investigate the beneficial effects of different antifreeze proteins (AFPs) on mouse ovarian tissue cryopreservation and transplantation.
Ovaries were obtained from 5-week-old B6D2F1 mice, and each ovary was cryopreserved using two-step vitrification and four-step warming procedures. In Experiment I, ovaries were randomly allocated into fresh, vitrification control, and nine experimental groups according to the AFP type (FfIBP, LeIBP, type III) and concentration (0.1, 1, 10 mg/mL) used. After vitrification and warming, 5,790 ovarian follicles were evaluated using histology and TUNEL assays, and immunofluorescence for τH2AX and Rad51 was used to detect DNA double-strand breaks (DSBs) and repair (DDR), respectively. In Experiment II, 20 mice were randomly divided into two groups: one where the vitrification and warming media were supplemented with 10 mg/mL LeIBP, and the other where media alone were used (control). Ovaries were then autotransplanted under both kidney capsules 7 days after vitrification together with the addition of 10 mg/mL LeIBP in the vitrification-warming media. After transplantation, the ovarian follicles, the percentage of apoptotic follicles, the extent of the CD31-positive area, and the serum FSH levels of the transplanted groups were compared.
In Experiment I, the percentage of total grade 1 follicles was significantly higher in the 10 mg/mL LeIBP group than in the vitrification control, while all AFP-treated groups had significantly improved grade 1 primordial follicle numbers compared with those of the vitrification control. The number of apoptotic (TUNEL-positive) follicles was significantly decreased in the groups treated with 1 and 10 mg/mL LeIBP. The proportion of τH2AX-positive follicles was significantly reduced in all AFP-treated groups, while the proportion of Rad51-positive follicles was significantly decreased in only the FfIBP- and LeIBP-treated groups. In Experiment II, after autotransplantation of OT vitrified with 10 mg/mL of LeIBP, the percentage of total grade 1 and primordial grade 1 follicles, and the extent of the CD31-positive area, were increased significantly. Moreover, the levels of serum FSH and the percentage of TUNEL-positive follicles were significantly lower in the LeIBP-treated than in the control group.
A supplementation with high concentrations of AFPs had protective effects on follicle preservation during OT vitrification-warming procedures. The group treated with LeIBP was protected most effectively. The beneficial effects of LeIBP were also observed after autotransplantation of vitrified-warmed OT. Further studies are necessary to determine the exact mechanism of these protective effects.</description><subject>Analysis</subject><subject>Animals</subject><subject>Antifreeze proteins</subject><subject>Antifreeze Proteins - metabolism</subject><subject>Antifreeze Proteins - pharmacology</subject><subject>Apoptosis</subject><subject>Apoptosis - drug effects</subject><subject>Cancer</subject><subject>Cancer therapies</subject><subject>Care and treatment</subject><subject>Complications and side effects</subject><subject>Cryopreservation</subject><subject>Cryopreservation - methods</subject><subject>Cryoprotective Agents - metabolism</subject><subject>Cryoprotective Agents - pharmacology</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA damage</subject><subject>Female</subject><subject>Fertility</subject><subject>Follicle Stimulating Hormone - blood</subject><subject>Follicle-stimulating hormone</subject><subject>Follicles</subject><subject>Gynecology</subject><subject>Health aspects</subject><subject>Histology</subject><subject>Immunofluorescence</subject><subject>Immunohistochemistry</subject><subject>Injury prevention</subject><subject>Kidneys</subject><subject>Leucosporidium</subject><subject>Medicine</subject><subject>Mice</subject><subject>Obstetrics</subject><subject>Ovarian cancer</subject><subject>Ovarian Follicle - cytology</subject><subject>Ovarian Follicle - drug effects</subject><subject>Ovaries</subject><subject>Ovary - cytology</subject><subject>Ovary - transplantation</subject><subject>Polypeptides</subject><subject>Proteins</subject><subject>Quality</subject><subject>Reproductive status</subject><subject>Supplements</subject><subject>Transplantation</subject><subject>Transplantation, Autologous</subject><subject>Vascular endothelial growth factor</subject><subject>Vitrification</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</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><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNk1-L1DAUxYso7jr6DUQLgujDjM3_9EVYllUHFhZ08TWk6e1Mlk5Tk3Rw_PRmZrrLVPZB-tBy7u-eNCe5WfYaFQtEBPp05wbf6XbRuw4WBcIcM_wkO0clwXOOC_L05PssexHCXVEwIjl_np1hVhJJKTvP4lXTgIkhd00e1x4gr21SPHQxj7seDgXdRduk2h_Ie-8i2C7JXb5xQ4DcbbW3usujDWGA3Pid6z0E8FsdbaJ0V-fR6y70bfI5aC-zZ41uA7wa37Ps9svV7eW3-fXN1-XlxfXc8BLHuaAV17wE2iBeCEaACcRrIII1GIsCG4qrkhjJiUFVyXEjKNQYeFNRUwhJZtnbo23fuqDGvIJCXEgmuKQsEcsjUTt9p3pvN9rvlNNWHQTnV0r7aE0LCuMakVJqqWtDa8G11sIgWlTSEKYrkbw-j6sN1QZqkxL0up2YTiudXauV2ypKkZSIJ4MPo4F3vwYIUW1sMNCm2CAlnf5bFkiyIp3qLHv3D_r47kZqpdMGbNe4tK7Zm6oLSvY-TJJELR6h0lPDxpp0uRqb9EnDx0lDYiL8jis9hKCWP77_P3vzc8q-P2HXoNu4Dq4d9jcmTEF6BI13IXhoHkJGhdrPxn0aaj8bapyN1Pbm9IAemu6HgfwFQyELcw</recordid><startdate>20150504</startdate><enddate>20150504</enddate><creator>Lee, Jaewang</creator><creator>Kim, Seul Ki</creator><creator>Youm, Hye Won</creator><creator>Kim, Hak Jun</creator><creator>Lee, Jung Ryeol</creator><creator>Suh, Chang Suk</creator><creator>Kim, Seok Hyun</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20150504</creationdate><title>Effects of three different types of antifreeze proteins on mouse ovarian tissue cryopreservation and transplantation</title><author>Lee, Jaewang ; Kim, Seul Ki ; Youm, Hye Won ; Kim, Hak Jun ; Lee, Jung Ryeol ; Suh, Chang Suk ; Kim, Seok Hyun</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-74b6a69e4f160753e5716de375f22702c42b93c863c1b962f74ed2e6fb4c0783</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Analysis</topic><topic>Animals</topic><topic>Antifreeze proteins</topic><topic>Antifreeze Proteins - metabolism</topic><topic>Antifreeze Proteins - pharmacology</topic><topic>Apoptosis</topic><topic>Apoptosis - drug effects</topic><topic>Cancer</topic><topic>Cancer therapies</topic><topic>Care and treatment</topic><topic>Complications and side effects</topic><topic>Cryopreservation</topic><topic>Cryopreservation - methods</topic><topic>Cryoprotective Agents - metabolism</topic><topic>Cryoprotective Agents - pharmacology</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA damage</topic><topic>Female</topic><topic>Fertility</topic><topic>Follicle Stimulating Hormone - blood</topic><topic>Follicle-stimulating hormone</topic><topic>Follicles</topic><topic>Gynecology</topic><topic>Health aspects</topic><topic>Histology</topic><topic>Immunofluorescence</topic><topic>Immunohistochemistry</topic><topic>Injury prevention</topic><topic>Kidneys</topic><topic>Leucosporidium</topic><topic>Medicine</topic><topic>Mice</topic><topic>Obstetrics</topic><topic>Ovarian cancer</topic><topic>Ovarian Follicle - cytology</topic><topic>Ovarian Follicle - drug effects</topic><topic>Ovaries</topic><topic>Ovary - cytology</topic><topic>Ovary - transplantation</topic><topic>Polypeptides</topic><topic>Proteins</topic><topic>Quality</topic><topic>Reproductive status</topic><topic>Supplements</topic><topic>Transplantation</topic><topic>Transplantation, Autologous</topic><topic>Vascular endothelial growth factor</topic><topic>Vitrification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lee, Jaewang</creatorcontrib><creatorcontrib>Kim, Seul Ki</creatorcontrib><creatorcontrib>Youm, Hye Won</creatorcontrib><creatorcontrib>Kim, Hak Jun</creatorcontrib><creatorcontrib>Lee, Jung Ryeol</creatorcontrib><creatorcontrib>Suh, Chang Suk</creatorcontrib><creatorcontrib>Kim, Seok Hyun</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lee, Jaewang</au><au>Kim, Seul Ki</au><au>Youm, Hye Won</au><au>Kim, Hak Jun</au><au>Lee, Jung Ryeol</au><au>Suh, Chang Suk</au><au>Kim, Seok Hyun</au><au>Schlatt, Stefan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effects of three different types of antifreeze proteins on mouse ovarian tissue cryopreservation and transplantation</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2015-05-04</date><risdate>2015</risdate><volume>10</volume><issue>5</issue><spage>e0126252</spage><epage>e0126252</epage><pages>e0126252-e0126252</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Ovarian tissue (OT) cryopreservation is effective in preserving fertility in cancer patients who have concerns about fertility loss due to cancer treatment. However, the damage incurred at different steps during the cryopreservation procedure may cause follicular depletion; hence, preventing chilling injury would help maintain ovarian function.
This study was designed to investigate the beneficial effects of different antifreeze proteins (AFPs) on mouse ovarian tissue cryopreservation and transplantation.
Ovaries were obtained from 5-week-old B6D2F1 mice, and each ovary was cryopreserved using two-step vitrification and four-step warming procedures. In Experiment I, ovaries were randomly allocated into fresh, vitrification control, and nine experimental groups according to the AFP type (FfIBP, LeIBP, type III) and concentration (0.1, 1, 10 mg/mL) used. After vitrification and warming, 5,790 ovarian follicles were evaluated using histology and TUNEL assays, and immunofluorescence for τH2AX and Rad51 was used to detect DNA double-strand breaks (DSBs) and repair (DDR), respectively. In Experiment II, 20 mice were randomly divided into two groups: one where the vitrification and warming media were supplemented with 10 mg/mL LeIBP, and the other where media alone were used (control). Ovaries were then autotransplanted under both kidney capsules 7 days after vitrification together with the addition of 10 mg/mL LeIBP in the vitrification-warming media. After transplantation, the ovarian follicles, the percentage of apoptotic follicles, the extent of the CD31-positive area, and the serum FSH levels of the transplanted groups were compared.
In Experiment I, the percentage of total grade 1 follicles was significantly higher in the 10 mg/mL LeIBP group than in the vitrification control, while all AFP-treated groups had significantly improved grade 1 primordial follicle numbers compared with those of the vitrification control. The number of apoptotic (TUNEL-positive) follicles was significantly decreased in the groups treated with 1 and 10 mg/mL LeIBP. The proportion of τH2AX-positive follicles was significantly reduced in all AFP-treated groups, while the proportion of Rad51-positive follicles was significantly decreased in only the FfIBP- and LeIBP-treated groups. In Experiment II, after autotransplantation of OT vitrified with 10 mg/mL of LeIBP, the percentage of total grade 1 and primordial grade 1 follicles, and the extent of the CD31-positive area, were increased significantly. Moreover, the levels of serum FSH and the percentage of TUNEL-positive follicles were significantly lower in the LeIBP-treated than in the control group.
A supplementation with high concentrations of AFPs had protective effects on follicle preservation during OT vitrification-warming procedures. The group treated with LeIBP was protected most effectively. The beneficial effects of LeIBP were also observed after autotransplantation of vitrified-warmed OT. Further studies are necessary to determine the exact mechanism of these protective effects.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>25938445</pmid><doi>10.1371/journal.pone.0126252</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2015-05, Vol.10 (5), p.e0126252-e0126252 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1678576845 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Analysis Animals Antifreeze proteins Antifreeze Proteins - metabolism Antifreeze Proteins - pharmacology Apoptosis Apoptosis - drug effects Cancer Cancer therapies Care and treatment Complications and side effects Cryopreservation Cryopreservation - methods Cryoprotective Agents - metabolism Cryoprotective Agents - pharmacology Deoxyribonucleic acid DNA DNA damage Female Fertility Follicle Stimulating Hormone - blood Follicle-stimulating hormone Follicles Gynecology Health aspects Histology Immunofluorescence Immunohistochemistry Injury prevention Kidneys Leucosporidium Medicine Mice Obstetrics Ovarian cancer Ovarian Follicle - cytology Ovarian Follicle - drug effects Ovaries Ovary - cytology Ovary - transplantation Polypeptides Proteins Quality Reproductive status Supplements Transplantation Transplantation, Autologous Vascular endothelial growth factor Vitrification |
| title | Effects of three different types of antifreeze proteins on mouse ovarian tissue cryopreservation and transplantation |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-26T06%3A44%3A17IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Effects%20of%20three%20different%20types%20of%20antifreeze%20proteins%20on%20mouse%20ovarian%20tissue%20cryopreservation%20and%20transplantation&rft.jtitle=PloS%20one&rft.au=Lee,%20Jaewang&rft.date=2015-05-04&rft.volume=10&rft.issue=5&rft.spage=e0126252&rft.epage=e0126252&rft.pages=e0126252-e0126252&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0126252&rft_dat=%3Cgale_plos_%3EA430193583%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1678576845&rft_id=info:pmid/25938445&rft_galeid=A430193583&rft_doaj_id=oai_doaj_org_article_22d1398a8adc4d76aaa7c140b8c35ab7&rfr_iscdi=true |