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http://dx.doi.org/10.5536/KJPS.2013.40.3.197

Effect of Ethylene Glycol(EG) and Propylene Glycol(PG) on the Viability of Frozen-thawed Primordial Germ Cells(PGCs) on Korean Native Chicken(Ogye) by Vitrification  

Kim, Hyun (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Kim, Dong Hun (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)
Ko, Yeoung Gyu (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Do, Yoon Jung (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.40, no.3, 2013 , pp. 197-205 More about this Journal
Abstract
This study established a method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for preservation of the species. The purpose of this study was to compare the effects of Ethylene Glycol (EG) and Propylene Glycol (PG) on viability of cryopreserved PGCs with vitrification in Korean Native Chicken (Ogye), and to fine should be find or to the optimal protocol for PGCs freezing. One of the important components of cryopreservation process is cryopreservation medium that plays a vital role in preventing cellular injury during freeze-thawing. Cryoprotective agents have been known to improve cell viability after freeze-thawing. PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents. Gonads were harvested from stage 28 chick embryos and pooled in groups of 10E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments: 2.5% EG, 5% EG, 10% EG, 2.5% PG, 5% PG, 10% PG, and 0% cryoprotectant as a control. Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. After freezing and thawing, survival rates of the frozen-thawed PGCs from the 0, 2.5, 5, 10 and 15% EG plus FBS treatment were 44.24%, 64.51%, 85.63%, 80.51% and 73.52% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05)(85.63% vs 66.81%). 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 PGC that enables systematic storage and labeling of cryopreserved PGCs in liquid N at a germplasm repository and ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.
Keywords
primordial germ cells (PGCs); vitrification; ethylene glycol (EG); propylene glycol (PG); viability; Korean Native Chicken (Ogye);
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Times Cited By KSCI : 2  (Citation Analysis)
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