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http://dx.doi.org/10.5536/KJPS.2014.41.4.261

The Effect of Simple Freezing Method on Viability of Frozen-thawed Primordial Germ Cells on the Chicken  

Kim, Hyun (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Cho, Young Moo (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Han, Jae Yong (WCU Biomodulation Major, Department of Agricultural Biotechnology, Seoul National University)
Choi, Sung Bok (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Cho, Chang-Yeon (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Suh, Sangwon (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Ko, Yeoung-Gyu (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Seong, Hwan-Hoo (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Kim, Sung Woo (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Publication Information
Korean Journal of Poultry Science / v.41, no.4, 2014 , pp. 261-270 More about this Journal
Abstract
This study was conducted to establish the method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of simple freeze-thaw treatment on viability of PGCs in chickens and to the optimal protocol for PGCs freezing. PGCs obtained from the germinal gonade of an early embryos of 5.5~6 day (stage 28) of Isa Brown, Korean Ogye (KO), White Leghorn and Commercial breeds, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15%, and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to simple freezing, with different concentrations of the cryoprotectant solution, were examined. After simple freezing, the viability of PGCs after freeze-thawing was significantly higher for Commercial breeds ($88.7{\pm}2.4%$) than KO ($85.1{\pm}0.4%$), Isa Brown ($84.6{\pm}0.2%$) and White Leghorn ($85.9{\pm}0.1%$) (p<0.05) using 10% EG cryoprotectant. Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at a germplasm repository and ease of entry into a database.
Keywords
primordial germ cells (PGCs); simple freezing; cryoprotective agents; viability; chicken breed;
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Times Cited By KSCI : 2  (Citation Analysis)
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