Document Detail


Feasibility of cryopreservation of zebrafish (Danio rerio) primordial germ cells by whole embryo freezing.
MedLine Citation:
PMID:  19827747     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Shogo Higaki; Kentaro Mochizuki; Hiroko Baba; Yuichiro Akashi; Etsuro Yamaha; Seiji Katagiri; Yoshiyuki Takahashi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Japanese journal of veterinary research     Volume:  57     ISSN:  0047-1917     ISO Abbreviation:  Jpn. J. Vet. Res.     Publication Date:  2009 Aug 
Date Detail:
Created Date:  2009-10-15     Completed Date:  2009-11-05     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0376567     Medline TA:  Jpn J Vet Res     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  119-28     Citation Subset:  IM    
Affiliation:
Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blastula / cytology
Butylene Glycols
Cell Division
Cryopreservation / methods
Cryoprotective Agents
Dimethyl Sulfoxide
Embryo, Nonmammalian / cytology*
Ethylene Glycol
Gastrula / cytology
Germ Cells / cytology*
Propylene Glycol
Zebrafish / embryology,  genetics*
Chemical
Reg. No./Substance:
0/Butylene Glycols; 0/Cryoprotective Agents; 107-21-1/Ethylene Glycol; 107-88-0/1,3-butylene glycol; 57-55-6/Propylene Glycol; 67-68-5/Dimethyl Sulfoxide

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