High in vitro development after somatic cell nuclear transfer and trichostatin A treatment of reconstructed porcine embryos
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High in vitro development after somatic cell nuclear transfer and trichostatin A treatment of reconstructed porcine embryos. / Li, J.; Østrup, Olga; Villemoes, Klaus; Kragh, Peter Michael; Schmidt, Mette; Bøgh, Ingrid Brück; Zhang, Y.; Du, Yutao; Lin, L.; Purup, Stig; Xue, Q.; Bolund, Lars; Yang, H.; Maddox-Hyttel, Poul; Vajta, Gabor.
In: Theriogenology, Vol. 70, No. 5, 2008, p. 800-808.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - High in vitro development after somatic cell nuclear transfer and trichostatin A treatment of reconstructed porcine embryos
AU - Li, J.
AU - Østrup, Olga
AU - Villemoes, Klaus
AU - Kragh, Peter Michael
AU - Schmidt, Mette
AU - Bøgh, Ingrid Brück
AU - Zhang, Y.
AU - Du, Yutao
AU - Lin, L.
AU - Purup, Stig
AU - Xue, Q.
AU - Bolund, Lars
AU - Yang, H.
AU - Maddox-Hyttel, Poul
AU - Vajta, Gabor
PY - 2008
Y1 - 2008
N2 - Abnormal epigenetic modification is supposed to be one of factors accounting for inefficient reprogramming of the donor cell nuclei in ooplasm after somatic cell nuclear transfer (SCNT). Trichostatin A (TSA) is an inhibitor of histone deacetylase, potentially enhancing cloning efficiency. The aim of our present study was to establish the optimal TSA treatment in order to improve the development of handmade cloned (HMC) porcine embryos and examine the effect of TSA on their development. The blastocyst percentage of HMC embryos treated with 37.5 nM TSA for 22-24 h after activation increased up to 80% (control grpup-54%; P<0.05). TSA mediated increase in histone acetylation was proved by immunofluorescence analysis of acH3K9 and acH4K16. 2-cell stage embryos derived from TSA treatment displayed significant increase in histone acetylation compared to control embryos, whereas no significant differences were observed at blastocyst stage. During time-lapse monitoring, no difference was observed in the kinetics of 2-cell stage embryos. Compact morula (CM) stage was reached 15 h later in TSA treated embryos compared to the control. Blastocysts (Day 5 and 6) from HMC embryos treated with TSA were transferred to 2 recipients resulting in one pregnancy and birth of one live and five dead piglets. Our data demonstrate that TSA treatment after HMC in pigs may affect reprogramming of the somatic genome resulting in higher in vitro embryo development, and enable full-term in vivo development.
AB - Abnormal epigenetic modification is supposed to be one of factors accounting for inefficient reprogramming of the donor cell nuclei in ooplasm after somatic cell nuclear transfer (SCNT). Trichostatin A (TSA) is an inhibitor of histone deacetylase, potentially enhancing cloning efficiency. The aim of our present study was to establish the optimal TSA treatment in order to improve the development of handmade cloned (HMC) porcine embryos and examine the effect of TSA on their development. The blastocyst percentage of HMC embryos treated with 37.5 nM TSA for 22-24 h after activation increased up to 80% (control grpup-54%; P<0.05). TSA mediated increase in histone acetylation was proved by immunofluorescence analysis of acH3K9 and acH4K16. 2-cell stage embryos derived from TSA treatment displayed significant increase in histone acetylation compared to control embryos, whereas no significant differences were observed at blastocyst stage. During time-lapse monitoring, no difference was observed in the kinetics of 2-cell stage embryos. Compact morula (CM) stage was reached 15 h later in TSA treated embryos compared to the control. Blastocysts (Day 5 and 6) from HMC embryos treated with TSA were transferred to 2 recipients resulting in one pregnancy and birth of one live and five dead piglets. Our data demonstrate that TSA treatment after HMC in pigs may affect reprogramming of the somatic genome resulting in higher in vitro embryo development, and enable full-term in vivo development.
KW - Former LIFE faculty
KW - Pig
KW - Handmade cloning
KW - Histone acetylation
KW - Time-lapse
KW - Reprogramming
U2 - 10.1016/j.theriogenology.2008.05.046
DO - 10.1016/j.theriogenology.2008.05.046
M3 - Journal article
VL - 70
SP - 800
EP - 808
JO - Theriogenology
JF - Theriogenology
SN - 0093-691X
IS - 5
ER -
ID: 8109737