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http://dx.doi.org/10.5536/KJPS.2014.41.4.249

The Evaluation of Various Conditions in the Cryopreservation of Primordial Germ Cells on Korean Native Chicken (Ogye)  

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)
Byun, Mi Jeong (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
Kim, Young Sin (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. 249-259 More about this Journal
Abstract
Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps and freezing media on the rates of viability of cryopreserved chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos of Korean Ogye (KO) and Commercial breeds (C), 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)). Gonads were harvested from stage 28 chick embryos and pooled in groups of 5, 10, 15, 20E embryos, contributing gonads to the cell suspension. 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 slow freezing and vitrification, with different concentrations of the cryoprotectant solution, were examined. After vitrification and slow freezing, survival rates of the frozen-thawed PGCs from the 10% EG plus FBS treatment were 85.63%, and 66.14% (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%) by vitrification. 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 data base. 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; slow freezing; viability; Korean native chicken (Ogye);
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