Cryopreservation of rat embryos at all developmental stages by small-volume vitrification procedure and rapid warming in cryotubes
Abstract Intracellular ice formation during cryopreservation is lethal to the cell, including during warming. Here, we examined the effect of sample volume and warming rate on the cryopreservation success of 1-cell rat embryos based on successful development into blastocysts in vitro and to term in...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-11-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-47394-0 |
| _version_ | 1797415571789185024 |
|---|---|
| author | Shinsuke Seki Toshiaki Kawabe Wataru Yamazaki Kazuaki Matsumura Takanori Oikawa Takahiro Obata Misako Higashiya Megumi Yano Tomoo Eto |
| author_facet | Shinsuke Seki Toshiaki Kawabe Wataru Yamazaki Kazuaki Matsumura Takanori Oikawa Takahiro Obata Misako Higashiya Megumi Yano Tomoo Eto |
| author_sort | Shinsuke Seki |
| collection | DOAJ |
| description | Abstract Intracellular ice formation during cryopreservation is lethal to the cell, including during warming. Here, we examined the effect of sample volume and warming rate on the cryopreservation success of 1-cell rat embryos based on successful development into blastocysts in vitro and to term in vivo following embryo transfer. Embryos were equilibrated in 5% propylene glycol solution for 10 min, held for 40 s at 0 °C in cryopreservation solution (5%PG + PEPeS), and cooled by immersion in liquid nitrogen. When 1-cell embryos were cryopreserved in a volume of 30–100 μL at a cooling rate of 5830–7160 °C/min and warmed at 35,480–49,400 °C/min by adding 1 mL of 0.3 M sucrose solution at 50 °C, 17.3–28.8% developed into blastocysts, compared with 57.0% of untreated embryos. However, when 1-cell embryos were cryopreserved in a smaller volume of 15 μl at 7950 °C/min and warmed at 68,850 °C/min, 58.8 ± 10.6% developed into blastocysts and 50.0 ± 7.4% developed to term, comparable to that of non-treated embryos (57.0 ± 5.4% and 51.4 ± 3.1%, respectively). Cryopreserved embryos at other developmental stages also showed high in vitro culture potential similar to that of the control. Using a conventional cryotube and a small-volume vitrification procedure with rapid warming, we achieved high levels of subsequent rat embryonic development at all developmental stages. |
| first_indexed | 2024-03-09T05:50:40Z |
| format | Article |
| id | doaj.art-c9e6afca0f9145ac9117681d1679d175 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-09T05:50:40Z |
| publishDate | 2023-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-c9e6afca0f9145ac9117681d1679d1752023-12-03T12:18:06ZengNature PortfolioScientific Reports2045-23222023-11-0113111010.1038/s41598-023-47394-0Cryopreservation of rat embryos at all developmental stages by small-volume vitrification procedure and rapid warming in cryotubesShinsuke Seki0Toshiaki Kawabe1Wataru Yamazaki2Kazuaki Matsumura3Takanori Oikawa4Takahiro Obata5Misako Higashiya6Megumi Yano7Tomoo Eto8Experimental Animal Division, Bioscience Education and Research Support Center, Akita UniversityARK Resource Co., Ltd.Experimental Animal Division, Bioscience Education and Research Support Center, Akita UniversitySchool of Materials Science, Japan Advanced Institute of Science and TechnologyExperimental Animal Division, Bioscience Education and Research Support Center, Akita UniversityExperimental Animal Division, Bioscience Education and Research Support Center, Akita UniversityExperimental Animal Division, Bioscience Education and Research Support Center, Akita UniversityExperimental Animal Division, Bioscience Education and Research Support Center, Akita UniversityCentral Institute for Experimental AnimalsAbstract Intracellular ice formation during cryopreservation is lethal to the cell, including during warming. Here, we examined the effect of sample volume and warming rate on the cryopreservation success of 1-cell rat embryos based on successful development into blastocysts in vitro and to term in vivo following embryo transfer. Embryos were equilibrated in 5% propylene glycol solution for 10 min, held for 40 s at 0 °C in cryopreservation solution (5%PG + PEPeS), and cooled by immersion in liquid nitrogen. When 1-cell embryos were cryopreserved in a volume of 30–100 μL at a cooling rate of 5830–7160 °C/min and warmed at 35,480–49,400 °C/min by adding 1 mL of 0.3 M sucrose solution at 50 °C, 17.3–28.8% developed into blastocysts, compared with 57.0% of untreated embryos. However, when 1-cell embryos were cryopreserved in a smaller volume of 15 μl at 7950 °C/min and warmed at 68,850 °C/min, 58.8 ± 10.6% developed into blastocysts and 50.0 ± 7.4% developed to term, comparable to that of non-treated embryos (57.0 ± 5.4% and 51.4 ± 3.1%, respectively). Cryopreserved embryos at other developmental stages also showed high in vitro culture potential similar to that of the control. Using a conventional cryotube and a small-volume vitrification procedure with rapid warming, we achieved high levels of subsequent rat embryonic development at all developmental stages.https://doi.org/10.1038/s41598-023-47394-0 |
| spellingShingle | Shinsuke Seki Toshiaki Kawabe Wataru Yamazaki Kazuaki Matsumura Takanori Oikawa Takahiro Obata Misako Higashiya Megumi Yano Tomoo Eto Cryopreservation of rat embryos at all developmental stages by small-volume vitrification procedure and rapid warming in cryotubes Scientific Reports |
| title | Cryopreservation of rat embryos at all developmental stages by small-volume vitrification procedure and rapid warming in cryotubes |
| title_full | Cryopreservation of rat embryos at all developmental stages by small-volume vitrification procedure and rapid warming in cryotubes |
| title_fullStr | Cryopreservation of rat embryos at all developmental stages by small-volume vitrification procedure and rapid warming in cryotubes |
| title_full_unstemmed | Cryopreservation of rat embryos at all developmental stages by small-volume vitrification procedure and rapid warming in cryotubes |
| title_short | Cryopreservation of rat embryos at all developmental stages by small-volume vitrification procedure and rapid warming in cryotubes |
| title_sort | cryopreservation of rat embryos at all developmental stages by small volume vitrification procedure and rapid warming in cryotubes |
| url | https://doi.org/10.1038/s41598-023-47394-0 |
| work_keys_str_mv | AT shinsukeseki cryopreservationofratembryosatalldevelopmentalstagesbysmallvolumevitrificationprocedureandrapidwarmingincryotubes AT toshiakikawabe cryopreservationofratembryosatalldevelopmentalstagesbysmallvolumevitrificationprocedureandrapidwarmingincryotubes AT wataruyamazaki cryopreservationofratembryosatalldevelopmentalstagesbysmallvolumevitrificationprocedureandrapidwarmingincryotubes AT kazuakimatsumura cryopreservationofratembryosatalldevelopmentalstagesbysmallvolumevitrificationprocedureandrapidwarmingincryotubes AT takanorioikawa cryopreservationofratembryosatalldevelopmentalstagesbysmallvolumevitrificationprocedureandrapidwarmingincryotubes AT takahiroobata cryopreservationofratembryosatalldevelopmentalstagesbysmallvolumevitrificationprocedureandrapidwarmingincryotubes AT misakohigashiya cryopreservationofratembryosatalldevelopmentalstagesbysmallvolumevitrificationprocedureandrapidwarmingincryotubes AT megumiyano cryopreservationofratembryosatalldevelopmentalstagesbysmallvolumevitrificationprocedureandrapidwarmingincryotubes AT tomooeto cryopreservationofratembryosatalldevelopmentalstagesbysmallvolumevitrificationprocedureandrapidwarmingincryotubes |