Feasibility of cryopreservation of zebrafish (Danio rerio) primordial germ cells by whole embryo freezing
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概要
- 論文の詳細を見る
We investigated the feasibility of cryopreservation of zebrafish (Danio rerio) blastomeres and primordial germ cells (PGCs) by rapid freezing of dechorionated whole embryos at the blastula, gastrula and segmentation stages. Initially we examined the glass-forming properties and embryo toxicities of 5 cryoprotectants: methanol (MeOH), ethylene glycol (EG), dimethyl sulfoxide (DMSO), propylene glycol (PG), and 1,3-butylene glycol (1,3-BG). Embryos at the blastula and gastrula stages had high sensitivities to cryoprotectant toxicities and were fragile against mechanical damage. Thus the segmentation stage embryos, the PGCs of which were visualized by injecting green fluorescence protein-nos1 3'UTR mRNA, were frozen using solutions containing each cryoprotectant at 6 M (first trial) and 2 types of cryoprotectants at 3 M each (second trial). In the first trial, live PGCs were recovered from most of the embryos frozen with EG (about 2 cells/embryo); however, a few embryos had live PGCs when embryos were frozen with other cryoprotectants. In the second trial, a mixture of EG + PG better preserved the viability of PGCs in frozen embryos. Live PGCs were recovered from all embryos frozen with EG + PG (about 3 cells/embryo), and the survival rate of PGCs was estimated to be about 25% based on the number of live PGCs in fresh embryos (about 12 cells/embryo). The present study indicates that we can utilize rapid freezing of dechorionated whole embryos at the segmentation stage for the cryopreservation of PGCs.
- Graduate School of Veterinary Medicine, Hokkaido Universityの論文
著者
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MOCHIZUKI Kentaro
Nanae Fresh-Water Laboratory, Field Science Center for Northern Biosphere, Hokkaido University
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Takahashi Yoshiyuki
北海道大学 獣医学研究科繁殖学教室
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Yamaha Etsuro
Nanae Fresh-water Laboratory Field Science Center For Northern Biosphere Hokkaido University
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Akashi Yuichiro
Laboratory Of Theriogenology Department Of Veterinary Clinical Sciences Graduate School Of Veterinar
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Takahashi Yoshiyuki
Graduate School Of Veterinary Medicine Hokkaido University
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Katagiri Seiji
Lab. Of Theriogenology Dep. Of Veterinary Clinical Sciences Graduate School Of Veterinary Medicine H
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Katagiri Seiji
Laboratory Of Theriogenology Department Of Clinical Sciences Graduate School Of Veterinary Medicine
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Higaki Shogo
Laboratory Of Theriogenology Department Of Veterinary Clinical Sciences Graduate School Of Veterinar
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Takahashi Y
Laboratory Of Theriogenology Department Of Veterinary Clinical Science Graduate School Of Veterinary
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Takahashi Yoshiyuki
Lab. Of Theriogenology Dep. Of Veterinary Clinical Sciences Graduate School Of Veterinary Medicine H
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Mochizuki Kentaro
Nanae Fresh-water Laboratory Field Science Center For Northern Biosphere Hokkaido University
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Takahashi Yoshiyuki
Laboratory Of Theriogenology Department Of Veterinary Clinical Sciences Graduate School Of Veterinar
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Takahashi Y
Laboratory Of Theriogenology Department Of Veterinary Clinical Sciences Graduate School Of Veterinar
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Takahashi Yoshiyuki
Laboratory Of Theriogenology Department Of Veterinary Clinical Sciences Graduate School Of Veterinary Medicine Hokkaido University
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