title: Derivation and characterization of putative embryonic stem cells from cloned rabbit embryos.
abstract: The present study aimed to establish embryonic stem (ES) cell lines, i.e., ntES cells, using rabbit blastocyst stage embryos cloned by somatic cell nuclear transfer. First, we investigated the development of cloned rabbit embryos reconstructed with normal fibroblasts and fibroblasts transfected with enhanced green fluorescence protein (eGFP). Blastocyst rates were 27.4% and 23.9%, respectively, for the embryos reconstructed with normal fibroblasts and fibroblasts transfected with eGFP compared with that from the parthenogenetic group (43.1%). One ntES cell line was established from embryos reconstructed with eGFP-transfected fibroblasts (1 of 17, 5.9%), and three ntES cell lines were derived from those with normal fibroblasts (3 of 17, 17.6%). All the ntES cell lines retained alkaline phosphatase activity and expressed ES cell-specific markers SSEA-4, Oct-4, TRA-1-60, and TRA-1-81. The pluripotency was further confirmed by reverse transcription-polymerase chain reaction analyses of Oct-4, Nanog, and Sox-2 expressions in ntES cell lines. The differentiation capacity of ntES cells was also examined in vitro and in vivo, by which these ntES cell lines were able to differentiate into all three germ layers through embryoid bodies and teratomas. In conclusion, it is apparent that the efficiency of ntES cells derived using eGFP-transfected donor cells is lower than that with nontransfected, normal fibroblasts donor cells. Similar to those from parthenogenetic embryos, all ntES cell lines derived from cloned rabbit embryos are able to express pluripotency markers and retain their capability to differentiate into various cell lineages both in vitro and in vivo.
authors.affiliation: Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, Republic of China; Division of Animal Science, School of Agriculture and Natural Resources, University of Phayao, Phayao, Thailand.
authors.forename: Payungsuk
authors.initials: P
authors.lastname: Intawicha
authors.affiliation: Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, Republic of China; Department of Animal Science, Faculty of Agriculture, Ubon Ratchathani University, Ubon Ratchathani, Thailand.
authors.forename: Chawalit
authors.initials: C
authors.lastname: Siriboon
authors.affiliation: Division of Biotechnology, Agriculture Technology Research Institute, Hsinchu City, Taiwan, Republic of China.
authors.forename: Chien-Hong
authors.initials: CH
authors.lastname: Chen
authors.affiliation: Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, Republic of China; Department of Medical Research, Taichung Veterans General Hospital, Taichung, Taiwan, Republic of China.
authors.forename: Yung-Tsung
authors.initials: YT
authors.lastname: Chiu
authors.affiliation: Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, Republic of China.
authors.forename: Tzu-An
authors.initials: TA
authors.lastname: Lin
authors.affiliation: Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, Republic of China; Departement d'Elevage, Institut du Developpement Rural, Universite Polytechnique de Bobo, Bobo Dioulasso, Burkina Faso.
authors.forename: Michel
authors.initials: M
authors.lastname: Kere
authors.affiliation: Department of Animal Science and Biotechnology, Tunghai University, Taichung, Taiwan, Republic of China.
authors.forename: Neng-Wen
authors.initials: NW
authors.lastname: Lo
authors.affiliation: Division of Biotechnology, Agriculture Technology Research Institute, Hsinchu City, Taiwan, Republic of China.
authors.forename: Kun-Hsiung
authors.initials: KH
authors.lastname: Lee
authors.affiliation: Department of Animal Science, Faculty of Agriculture, Ubon Ratchathani University, Ubon Ratchathani, Thailand; Department of Biomedical Informatics and Medical Engineering, Asia University, Taichung, Taiwan, Republic of China.
authors.forename: Li-Yung
authors.initials: LY
authors.lastname: Chang
authors.affiliation: Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, Republic of China.
authors.forename: Hsing-I
authors.initials: HI
authors.lastname: Chiang
authors.affiliation: Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, Republic of China; Department of Animal Science, Faculty of Agriculture, Ubon Ratchathani University, Ubon Ratchathani, Thailand; Department of Biomedical Informatics and Medical Engineering, Asia University, Taichung, Taiwan, Republic of China; Core Laboratory for Stem Cell Research, Medical Research Department, China Medical University Hospital, Taichung, Taiwan, Republic of China. Electronic address: jcju@dragon.nchu.edu.tw.
authors.forename: Jyh-Cherng
authors.initials: JC
authors.lastname: Ju
mesh_terms: D000818:Animals; D001755:Blastocyst; D019976:Cloning, Organism / Q000662:veterinary*; D004622:Embryo, Mammalian / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000502:physiology*; D059785:Karyotype; D011817:Rabbits / Q000196:embryology*
keywords: Embryoid bodies; Embryonic stem cells; Rabbit ES cells; Somatic cell nuclear transfer