title: Production of viable cloned miniature pigs by aggregation of handmade cloned embryos at the 4-cell stage.
abstract: The aim of the present study was to improve the quality of handmade cloned porcine embryos by multiple embryo aggregations. Embryos derived from aggregation of three cloned embryos (3×) had a better blastocyst rate than cloned control (1×) embryos (73.6% vs 35.1%, respectively; P<0.05), but did not differ from those produced by aggregation of two cloned embryos (2×; 63.0%). Total cell numbers differed among treatments (P<0.05), with the greatest cell numbers (126) in the 3× group and the lowest (55) in the control group. The ratio of inner cell mass:total cell number was comparable in the 2× and 3× groups (25.1% vs 26.1%, respectively) and was significantly better than that in the control group (15.3%). The proportion of apoptotic cells in 2× and 3× groups was lower than that in the control group (2.7% and 2.2% vs 4.7%, respectively; P<0.05). Expression of Oct4 and Cdx2 was higher, whereas that of Bax was lower (P<0.05), in the 3× compared with non-aggregate group. Seven piglets were born to two surrogate mothers after embryo transfer of 3× aggregated blastocysts. In conclusion, aggregated embryos had greater total cell numbers and better pluripotency gene expression, with reduced expression of the pro-apoptosis gene Bax. Collectively, these improvement may be associated with the development of cloned embryos to term.
authors.affiliation: Department of Animal Science, National Chung Hsing University, 250 Kuokuang Road, Taichung 402, Taiwan, ROC.
authors.forename: Chawalit
authors.initials: C
authors.lastname: Siriboon
authors.affiliation: Animal Technology Institute Taiwan, 52 Kedung 2 Road, Ding-Pu LII, Chunan, Miaoli, Taiwan, ROC.
authors.forename: Ching-Fu
authors.initials: CF
authors.lastname: Tu
authors.affiliation: Department of Animal Science, National Chung Hsing University, 250 Kuokuang Road, Taichung 402, Taiwan, ROC.
authors.forename: Michel
authors.initials: M
authors.lastname: Kere
authors.affiliation: Animal Technology Institute Taiwan, 52 Kedung 2 Road, Ding-Pu LII, Chunan, Miaoli, Taiwan, ROC.
authors.forename: Ming-Sing
authors.initials: MS
authors.lastname: Liu
authors.affiliation: Animal Technology Institute Taiwan, 52 Kedung 2 Road, Ding-Pu LII, Chunan, Miaoli, Taiwan, ROC.
authors.forename: Hui-Jung
authors.initials: HJ
authors.lastname: Chang
authors.affiliation: Animal Technology Institute Taiwan, 52 Kedung 2 Road, Ding-Pu LII, Chunan, Miaoli, Taiwan, ROC.
authors.forename: Lin-Lin
authors.initials: LL
authors.lastname: Ho
authors.affiliation: Animal Technology Institute Taiwan, 52 Kedung 2 Road, Ding-Pu LII, Chunan, Miaoli, Taiwan, ROC.
authors.forename: Miao-En
authors.initials: ME
authors.lastname: Tai
authors.affiliation: Animal Technology Institute Taiwan, 52 Kedung 2 Road, Ding-Pu LII, Chunan, Miaoli, Taiwan, ROC.
authors.forename: Wen-Der
authors.initials: WD
authors.lastname: Fang
authors.affiliation: Department of Animal Science and Biotechnology, Tunghai University, 181, Sec. 3, Taichung Harbor Road, Taichung 407, Taiwan, ROC.
authors.forename: Neng-Wen
authors.initials: NW
authors.lastname: Lo
authors.affiliation: School of Optometry, Chung Shan Medical University, 110 Chien-Kuo North Road, Taichung 402, Taiwan, ROC.
authors.forename: Jung-Kai
authors.initials: JK
authors.lastname: Tseng
authors.affiliation: Department of Animal Science, National Chung Hsing University, 250 Kuokuang Road, Taichung 402, Taiwan, ROC.
authors.forename: Jyh-Cherng
authors.initials: JC
authors.lastname: Ju
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D053624:Blastocyst Inner Cell Mass / Q000166:cytology; D000071616:CDX2 Transcription Factor; D002449:Cell Aggregation / Q000502:physiology*; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D017931:DNA Primers / Q000235:genetics; D004622:Embryo, Mammalian / Q000196:embryology*; D005260:Female; D020869:Gene Expression Profiling; D018398:Homeodomain Proteins / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D011247:Pregnancy; D011256:Pregnancy Outcome; D060888:Real-Time Polymerase Chain Reaction; D013552:Swine; D013556:Swine, Miniature / Q000196:embryology*; D015534:Trans-Activators / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000071616:CDX2 Transcription Factor; D017931:DNA Primers; D018398:Homeodomain Proteins; D050814:Octamer Transcription Factor-3; D015534:Trans-Activators; D051028:bcl-2-Associated X Protein