title: Isolation of chicken embryonic stem cell and preparation of chicken chimeric model.
abstract: Chicken embryonic stem cells (ESCs) were separated from blastoderms at stage-X and cultured in vitro. Alkaline phosphatase activity and stage-specific embryonic antigen-1 staining was conducted to detect ESCs. Then, chicken ESCs were transfected with linearized plasmid pEGFP-N1 in order to produce chimeric chicken. Firstly, the optimal electrotransfection condition was compared; the results showed the highest transfection efficiency was obtained when the field strength and pulse duration was 280 V and 75 μs, respectively. Secondly, the hatchability of shedding methods, drilling a window at the blunt end of egg and drilling a window at the lateral shell of egg was compared, the results showed that the hatchability was the highest for drilling a window at the lateral shell of egg. Thirdly, the hatchability of microinjection (ESCs was microinjected into chick embryo cavity) was compared too, the results showed there were significant difference between the injection group transfected with ESCs and that of other two groups. In addition, five chimeric chickens were obtained in this study and EGFP gene was expressed in some organs, but only two chimeric chicken expressed EGFP gene in the gonad, indicating that the chimeric chicken could be obtained through chick embryo cavity injection by drilling a window at the lateral shell of egg.
authors.affiliation: College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
authors.forename: Yani
authors.initials: Y
authors.lastname: Zhang
authors.forename: Haiyan
authors.initials: H
authors.lastname: Yang
authors.forename: Zhentao
authors.initials: Z
authors.lastname: Zhang
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Shi
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Zheng
authors.forename: Bichun
authors.initials: B
authors.lastname: Li
authors.forename: Jiuzhou
authors.initials: J
authors.lastname: Song
mesh_terms: D000818:Animals; D001756:Blastoderm; D002470:Cell Survival; D002642:Chick Embryo; D002645:Chickens; D002678:Chimera / Q000196:embryology*; D053595:Embryonic Stem Cells / Q000166:cytology*; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics; D008845:Microinjections; D010957:Plasmids; D014162:Transfection
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins