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Volume 57, Number 2 >

Feasibility of cryopreservation of zebrafish (Danio rerio) primordial germ cells by whole embryo freezing

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Title: Feasibility of cryopreservation of zebrafish (Danio rerio) primordial germ cells by whole embryo freezing
Authors: Higaki, Shogo Browse this author
Mochizuki, Kentaro Browse this author
Baba, Hiroko Browse this author
Akashi, Yuichiro Browse this author
Yamaha, Etsuro Browse this author →KAKEN DB
Katagiri, Seiji Browse this author
Takahashi, Yoshiyuki Browse this author →KAKEN DB
Keywords: blastomere
primordial germ cell
Issue Date: Aug-2009
Publisher: Graduate School of Veterinary Medicine, Hokkaido University
Journal Title: Japanese Journal of Veterinary Research
Volume: 57
Issue: 2
Start Page: 119
End Page: 128
Abstract: 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.
Type: bulletin (article)
Appears in Collections:Japanese Journal of Veterinary Research > Volume 57, Number 2

Submitter: 獣医学部図書室

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