Int. J. Dev. Biol. 54: 1493 - 1501 (2010)
doi: 10.1387/ijdb.093059yk
© UPV/EHU Press

Generation of germ-line chimera zebrafish using primordial germ cells isolated from cultured blastomeres and cryopreserved embryoids

Yutaka Kawakami*,1, Rie Goto-Kazeto1, Taiju Saito1, Takafumi Fujimoto1, Shogo Higaki3, Yoshiyuki Takahashi2, Katsutoshi Arai3 and Etsuro Yamaha1

1Nanae Fresh Water Laboratory, Field Science Center of Northern Biosphere, Hokkaido University, Nanae, Japan, 2Laboratory of Theriogenology, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo and 3Laboratory of Aquaculture Genetics and Genomics, Graduate School of Fisheries Sciences, Hokkaido University, Hokkaido, Japan

ABSTRACT Primordial germ cells (PGCs) are the only cells in developing embryos with the potential to transmit genetic information to the next generation. In our previous study, a single PGC transplanted into a host differentiated into fertile gametes and produced germ-line chimeras of cyprinid fish, including zebrafish. In this study, we aimed to induce germ-line chimeras by transplanting donor PGCs from various sources (normal embryos at different stages, dissociated blastomeres, embryoids, or embryoids cryopreserved by vitrification) into host blastulae, and compare the migration rates of the PGCs towards the gonadal ridge. Isolated, cultured blastomeres not subject to mesodermal induction were able to differentiate into PGCs that retained their motility. Moreover, these PGCs successfully migrated towards the gonadal ridge of the host and formed viable gametes. Motility depended on developmental stage and culture duration: PGCs obtained at earlier developmental stages and with shorter cultivation periods showed an increased rate of migration to the gonadal ridge. Offspring were obtained from natural spawning between normal females and chimeric males. These results provide the basis for new methods of gene preservation in zebrafish.


germ-line chimera, primordial germ cell, cryopreservation, dissociated blastomere

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