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[Poster Presentation] - Cloning

The Effect of Reprogramming Donor Cells with Porcine iPSC Extracts on the In Vitro Development of Porcine Nuclear Transfer Embryos

초록

영어

Limited success of somatic cell nuclear transfer(SCNT) is attributed to incomplete reprogramming of transferred donor cell. Several approachs, such as histone deacetylase inhibitors and DNA methyltransferase inhibitors have been used to improve the efficiency of somatic cell nuclear transfer. Recently, it is reported that pre-treatment of somatic cells with undifferentiated cell extract, such as embryonic stem cell and mammalian oocytes is an attractive alternative ways to reprogramming control. The aim of this study was to evaluate the early development of porcine cloned embryos produced with porcine ear skin fibroblasts pre-treated with extract from porcine induced pluripotent stem cell (iPSC). For transport of porcine iPSC extract into cultured porcine ear skin fibroblasts, the ChariotTM reagent system was used. Treated cells were cultured for 3 days, and used for the analysis of histone H3K9 acetylation and SCNT The acetylation status of H3K9 was increased in cells treated with iPSC extract and cultured for 3 days compared with control. But, no significant difference was observed between the extract treated and control groups. After SCNT. no difference was observed in the rate of fusion (86.6% vs 86.2%) and embryo cleavage (86.6% vs 87.1%) between the extract treated and control groups. Also, no significant difference was noted in blastocyst rates (23.4% vs 28.4%) as well as cell numbers (43.8±10.8 vs 41.2±11.6) with extract treated group compared with control group. Overall apoptosis rate in blastocyst was not differences between the extract treated and control groups (4.6±3.5% vs 6.0± 5.8%). However, blastocyst rate with high apoptotic cells(>10% appototic cells) was significantly lower in extract treated group when compared with control group (7.1% vs 21.8%).. Our results demonstrated that pre-treatment of porcine ear skin fibroblasts using porcine iPSc extract had beneficial effect on the decreasing apoptosis in the blastocyst cultured in vitro, although there was no effect on the embryonic development.

저자정보

  • Jin-Gu No Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration
  • Mi-Kyung Choi Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration
  • Dae-Jin Kwon Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration
  • Jong-Ju Park Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration
  • Jea-Kyu Yoo Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration
  • Dong-Hoon Kim Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration

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