title: Derivation of Porcine Embryonic Stem-Like Cells from In Vitro-Produced Blastocyst-Stage Embryos.
abstract: Efficient isolation of embryonic stem (ES) cells from pre-implantation porcine embryos has remained a challenge. Here, we describe the derivation of porcine embryonic stem-like cells (pESLCs) by seeding the isolated inner cell mass (ICM) from in vitro-produced porcine blastocyst into α-MEM with basic fibroblast growth factor (bFGF). The pESL cells kept the normal karyotype and displayed flatten clones, similar in phenotype to human embryonic stem cells (hES cells) and rodent epiblast stem cells. These cells exhibited alkaline phosphatase (AP) activity and expressed pluripotency markers such as OCT4, NANOG, SOX2, SSEA-4, TRA-1-60, and TRA-1-81 as determined by both immunofluorescence and RT-PCR. Additionally, these cells formed embryoid body (EB), teratomas and also differentiated into 3 germ layers in vitro and in vivo. Microarray analysis showed the expression of the pluripotency markers, PODXL, REX1, SOX2, KLF5 and NR6A1, was significantly higher compared with porcine embryonic fibroblasts (PEF), but expression of OCT4, TBX3, REX1, LIN28A and DPPA5, was lower compared to the whole blastocysts or ICM of blastocyst. Our results showed that porcine embryonic stem-like cells can be established from in vitro-produced blastocyst-stage embryos, which promote porcine naive ES cells to be established.
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Dao-Rong
authors.initials: DR
authors.lastname: Hou
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Nie
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Man-Ling
authors.initials: ML
authors.lastname: Zhang
authors.affiliation: The Key Laboratory of the National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, Inner Mongolia, China.
authors.forename: Na
authors.initials: N
authors.lastname: Ta
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Li-Hua
authors.initials: LH
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Zhao-Qiang
authors.initials: ZQ
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Hai-Bin
authors.initials: HB
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Yan-Ru
authors.initials: YR
authors.lastname: Li
authors.affiliation: State Key Laboratory of Reproductive Biology; Institute of Zoology; Chinese Academy of Sciences, Beijing, China.
authors.forename: Qing-Yuan
authors.initials: QY
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Yi-Fan
authors.initials: YF
authors.lastname: Dai
authors.affiliation: State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China.|Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China.
authors.forename: Rong-Feng
authors.initials: RF
authors.lastname: Li
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001755:Blastocyst / Q000166:cytology*; D002454:Cell Differentiation; D048430:Cell Shape; D016000:Cluster Analysis; D003114:Colony-Forming Units Assay; D058732:Embryoid Bodies / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005307:Fertilization in Vitro; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D034421:Sus scrofa; D013724:Teratoma / Q000473:pathology; D014158:Transcription, Genetic
chemical_list: D015415:Biomarkers