title: Big Animal Cloning Using Transgenic Induced Pluripotent Stem Cells: A Case Study of Goat Transgenic Induced Pluripotent Stem Cells.
abstract: Using of embryonic stem cells (ESCs) could improve production traits and disease resistance by improving the efficiency of somatic cell nuclear transfer (SCNT) technology. However, robust ESCs have not been established from domestic ungulates. In the present study, we generated goat induced pluripotent stem cells (giPSCs) and transgenic cloned dairy goat induced pluripotent stem cells (tgiPSCs) from dairy goat fibroblasts (gFs) and transgenic cloned dairy goat fibroblasts (tgFs), respectively, using lentiviruses that contained hOCT4, hSOX2, hMYC, and hKLF4 without chemical compounds. The giPSCs and tgiPSCs expressed endogenous pluripotent markers, including OCT4, SOX2, MYC, KLF4, and NANOG. Moreover, they were able to maintain a normal karyotype and differentiate into derivatives from all three germ layers in vitro and in vivo. Using SCNT, tgFs and tgiPSCs were used as donor cells to produce embryos, which were named tgF-Embryos and tgiPSC-Embryos. The fusion rates and cleavage rates had no significant differences between tgF-Embryos and tgiPSC-Embryos. However, the expression of IGF-2, which is an important gene associated with embryonic development, was significantly lower in tgiPSC-Embryos than in tgF-Embryos and was not significantly different from vivo-Embryos.
authors.affiliation: 1 Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University , Nanjing, 210095, P.R. China .|2 Department of Medical Genetic and Cell Biology, Ningxia Medical University , Yinchuan, 750004, China .
authors.forename: Hui
authors.initials: H
authors.lastname: Song
authors.affiliation: 1 Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University , Nanjing, 210095, P.R. China .|2 Department of Medical Genetic and Cell Biology, Ningxia Medical University , Yinchuan, 750004, China .
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: 1 Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University , Nanjing, 210095, P.R. China .
authors.forename: Mingrui
authors.initials: M
authors.lastname: Huang
authors.affiliation: 3 Stanford University School of Medicine , Stanford, CA, 94305.
authors.forename: Dan
authors.initials: D
authors.lastname: Xu
authors.affiliation: 1 Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University , Nanjing, 210095, P.R. China .
authors.forename: Ziyu
authors.initials: Z
authors.lastname: Wang
authors.affiliation: 1 Jiangsu Engineering Technology Research Center of Meat Sheep & Goat Industry, Nanjing Agricultural University , Nanjing, 210095, P.R. China .
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods* / Q000662:veterinary; D003612:Dairying; D004423:Ear; D004622:Embryo, Mammalian; D005347:Fibroblasts / Q000166:cytology; D006041:Goats / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051379:Mice; D053652:Nuclear Transfer Techniques / Q000662:veterinary*