title: Efficiency of donor cell preparation and recipient oocyte source for production of transgenic cloned dairy goats harboring human lactoferrin.
abstract: The objective was to investigate the effects of the transgenic donor cell synchronization method, oocyte sources, and other factors, on production of hLF-gene nucleus transfer dairy goats. Three transfected cell lines from ear biopsies from three 3-mo-old Saanen dairy goats (designated Number 1, Number 2, and Number 3, respectively) were selected as karyoplast donors for somatic cell nuclear transfer (SCNT) after detailed identification (including PCR and sequencing of PCR products). In donor cell cycle synchronization studies, the apoptosis rate of hLF transgenic fibroblasts was not different (P > 0.05) after 3 days of serum starvation or 2 days of contact inhibition. Additionally, there was no effect (P > 0.05) on developmental capacity of reconstructed embryos; however, the kidding rate of recipients in the serum starvation group was higher than that in the contact inhibition group (18 vs. 0%, respectively). The production efficiency of the transgenic cloned goats using donor cells from the Number 1 dairy goat cell line was higher than those using the Number 2 and the Number 3 cell lines (kidding rates were 18, 2, and 0%, respectively, P < 0.05). The oocyte source did not significantly affect the pregnancy rate of hLF-transgenic cloned dairy goats, but more fetuses were aborted when using in vitro matured oocytes compared to in vivo matured oocytes. In summary, utilizing transfected 3-mo-old dairy goat fibroblasts as donor cells, seven live offspring were produced, and the hLF gene was successfully integrated. This study provided additional insights into preparation of donor cells and recipient oocytes for producing transgenic cloned goats through SCNT.
authors.affiliation: Center of Embryo Engineering and Technology, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Yong-Jie
authors.initials: YJ
authors.lastname: Wan
authors.forename: Yan-Li
authors.initials: YL
authors.lastname: Zhang
authors.forename: Zheng-Rong
authors.initials: ZR
authors.lastname: Zhou
authors.forename: Ruo-Xin
authors.initials: RX
authors.lastname: Jia
authors.forename: Meng
authors.initials: M
authors.lastname: Li
authors.forename: Hui
authors.initials: H
authors.lastname: Song
authors.forename: Zi-Yu
authors.initials: ZY
authors.lastname: Wang
authors.forename: Li-Zhong
authors.initials: LZ
authors.lastname: Wang
authors.forename: Guo-Min
authors.initials: GM
authors.lastname: Zhang
authors.forename: Ji-Hao
authors.initials: JH
authors.lastname: You
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D004624:Embryo Transfer / Q000662:veterinary; D005260:Female; D005347:Fibroblasts / Q000648:ultrastructure; D006041:Goats / Q000235:genetics*; D006801:Humans; D007781:Lactoferrin / Q000235:genetics*; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000254:growth & development / Q000648:ultrastructure; D011247:Pregnancy; D014162:Transfection
chemical_list: D007781:Lactoferrin