Earticle

초록

영어

Somatic cell nuclear transfer (SCNT) and induced pluripotent stem cell (iPS) experiments have generally demonstrated that a differentiated cell directly converts into a undifferentiated or pluripotent state. In SCNT experiment, nuclear reprogramming is induced by exposure of introduced donor cell nuclei to the recipient cytoplasm of matured oocytes. Although nuclear reprogramming of cells by the ex-ovo methods is not always consistent or efficient, it has been suggested that a combination of nuclear reprogramming technique may improve the efficiency or frequency of normal development of SCNT embryos. Here we hypothesized that treatment of somatic cells with extracts prepared from GV stage sturgeon's oocytes prior to their use as nuclear donors for SCNT will improve subsequent development. We reported a reversible permeabilization protocol with digitonin to deliver sturgeon oocyte exteact (SOE) to porcine fetal fibroblast cell nuclei ex ovo. Porcine fibroblasts were permeabilized by 4 μg/ml of digitonin for 2 min at 4℃ and then incubated in SOE for 7h at 15 18℃ followed by resealing of cell membrane. As results, no difference was observed in the number of fused couplets or the number of fused couplets that cleaved between the extract treated or control group. However, there was a significantly decrease in the percentage of fused couplets that developed to the blastocyst stage in the SOE treated group (p<0.05). Histone acetylation status was determined using an antibody to acetylation at lysine 9 on histone 3 (H3K9Ac). The intensity of H3K9Ac staining in 1-cell stage NT embryos was significantly increased when treated with the SOE (p<0.05), similar to that in 1-cell stage IVF embryos. In addition, porcine NT embryos reconstructed by using donor cell exposed to SOE prior to cell fusion significantly decreased developmental competence to the blastocyst stage but increased pluripotent gene expressions (Sox2, Nanog and Oct3/4) when compared with those in normal NT embryos (p<0.05).

키워드

저자

• So-Young Kim [ Department of Animal Biosciences, Gyeongsang National University ]
• Sang-Hoon Park [ Department of Animal Biosciences, Gyeongsang National University ]
• Mi-Ran Lee [ Department of Animal Biosciences, Gyeongsang National University ]
• Hye-Ju Eun [ Department of Animal Biosciences, Gyeongsang National University ]
• Sang-Ki Baik [ Department of Animal Biosciences, Gyeongsang National University ]
• Tae-Suk Kim [ Department of Animal Biosciences, Gyeongsang National University ]
• Joon-Hee Lee [ Department of Animal Biosciences, Gyeongsang National University ]

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간행물

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• 간행물명

  발생공학 국제심포지엄 및 학술대회 [International Symposium on Developmental Biotechnology]

• 간기

  연간

• 수록기간

  2004~2018

• 십진분류

  KDC 527 DDC 636