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- Title
Technical note: Viability and motility of vitrified/thawed primordial germ cell isolated from common carp (Cyprirtus carpio) somite embryos.
- Authors
Kawakami, Y.; Saito, T.; Fujimoto, T.; Goto-Kazeto, R.; Takahashi, E.; Adachi, S.; Arai, K.; Yamaha, E.
- Abstract
The feasibility of cryopreserving common carp (Cyprinus carpio) primordial germ cells (PGC) by vitrification of whole embryos at the 22to 28-somite stage was investigated. Green fluorescent protein (GFP)-labeled PGC were cooled rapidly using liquid nitrogen after exposure to a pretreatment solution containing 1.5 M cryoprotectant (ethylene glycol or dimethyl sulfoxide, 30 or 50 mm) and a vitrification solution containing 3 M cryoprotectant and 0.5 M sucrose (5, 10, 20, or 30 mm). Embryonic cells that were pretreated for 30 mill and vitrified for 20 mill with ethylene glycol had the greatest rate of survival of embryonic cells (68.6%; P < 0.01), an optimal highest percentage of viable POC (73.8 to 74.9%; P < 0.05), and no evidence of ice formation after thawing. The vitrified/thawed PGC were transplanted into blastulastage embryos from goldfish (Carassius auratus). The PGC maintained their motility and moved to the gonadal ridge of the host embryo. Thus, the combination of vitrification and transplantation to produce germline chimeras is a powerful tool for the artificial production of next-generation offspring.
- Subjects
CRYOPRESERVATION of organs, tissues, etc.; CARP; EMBRYOS; GERM cells; GOLDFISH; CHIMERISM
- Publication
Journal of Animal Science, 2012, Vol 90, Issue 2, p495
- ISSN
0021-8812
- Publication type
Article
- DOI
10.2527/jas.2011-4329