Document Detail

Production of fertile zebrafish (Danio rerio) possessing germ cells (gametes) originated from primordial germ cells recovered from vitrified embryos.
MedLine Citation:
PMID:  20154175     Owner:  NLM     Status:  MEDLINE    
This study aimed to produce fertile zebrafish (Danio rerio) possessing germ cells (gametes) that originated from cryopreserved primordial germ cells (PGCs). First, to improve the vitrification procedure of PGCs in segmentation stage embryos, dechorionated yolk-intact and yolk-removed embryos, the PGCs of which were labeled with green fluorescent protein, were cooled rapidly after serial exposures to equilibration solution (ES) and vitrification solution (VS), which contained ethylene glycol, DMSO, and sucrose. Yolk removal well prevented ice formation in the embryos during cooling and improved the viability of cryopreserved PGCs. The maximum recovery rate of live PGCs in the yolk-removed embryos vitrified after optimum exposure to ES and VS was estimated to be about 90%, and about 50% of the live PGCs showed pseudopodial movement. Next, to elucidate the ability of cryopreserved PGCs to differentiate into functional gametes, PGCs recovered from the yolk-removed embryos (striped-type) that were vitrified under the optimum exposure to ES and VS were transplanted individually into 218 sterilized recipient blastulae (golden-type). Two days after the transplantation, 7.5% (14/187) of morphologically normal embryos had PGC(s) in the genital ridges. Six (5 males and 1 female) of the 14 recipient embryos developed into mature fish and generated progeny with characteristics inherited from PGC donors. In conclusion, we demonstrated the successful cryopreservation of PGCs by vitrification of yolk-removed embryos and the production of fertile zebrafish possessing germ cells that originated from the PGCs in vitrified embryos.
Shogo Higaki; Yoshiki Eto; Yutaka Kawakami; Etsuro Yamaha; Noriko Kagawa; Masashige Kuwayama; Masashi Nagano; Seiji Katagiri; Yoshiyuki Takahashi
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-02-12
Journal Detail:
Title:  Reproduction (Cambridge, England)     Volume:  139     ISSN:  1741-7899     ISO Abbreviation:  Reproduction     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-03-23     Completed Date:  2010-07-05     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100966036     Medline TA:  Reproduction     Country:  England    
Other Details:
Languages:  eng     Pagination:  733-40     Citation Subset:  IM    
Laboratory of Theriogenology, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
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MeSH Terms
Cell Differentiation / physiology
Cell Separation
Cell Survival / physiology
Cells, Cultured
Embryo Culture Techniques
Embryo, Nonmammalian*
Fertility / physiology*
Germ Cells / cytology,  physiology*
Yolk Sac / cytology
Zebrafish / embryology*,  physiology

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