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KMID : 1023620120360010065
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Reproductive and Developmental Biology
2012 Volume.36 No. 1 p.65 ~ p.70
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The Effect of Modified Cryopreservation Method on Viability of Frozen-thawed Blastodermal Cells on the Korean Native Chicken(Ogolgye Breed)
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Kim Hyun
Kim Dong-Hoon
Park Soo-Bong
Choi Seong-Bok
Ko Yeoung-Gyu
Kim Jae-Hwan
Do Yoon-Jung
Park Hae-Geum
Kim Sung-Woo
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Abstract
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For reconstituting genetic resource(Korean Native Chicken: KNC) with grem-line chimeric chicken made with cryopreserved biastdermal cells, the experiments were carried out to optimize cryopreservating conditions. Stage X biastdemal cells were collected from KNC embryos and dissociated. Cells were susupended in medium containing cyopretectant and fetal bovine serum(FBS), and distributed into plastic ampules. Cell susupensions were seeded to induce ice formation at ?7 ℃ to ?35 ℃ at in the experiments, the effect of modification of dissociation way, concentration of FBS and cell density on the vaibility of frezen-thawed cells were investigated by trypan blue exclusion. Then change the way of cell dissociation from pipetting to short time vortexing, viability of frozen-thawed cell tended to be increaced from 29 % to 52 %. Increase concentraition of FBS in frozen medium from 20 % to 80 % made viability of thawed cell from 28 % to 35 %. The viability of thawed cells were 33.9% frozen at 2 embryos/ 0.5ml, and 43.6 % frozen at 20 embryos/0.5 ml. Furthermore, combination of three modifications make big improvement. The viability of frozen-thawed cell was 60 % for combinated method, and 41 % for general method. This result means the advance
to practical cryoreservation of blastdermal cell of the KNC(Ogolgye breed).
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KEYWORD
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Blastodermal cells, Cryopreservation method, Viability, Korean native chicken(Ogolgye breed)
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