title: Isolation and culture of embryonic stem-like cells from pig nuclear transfer blastocysts of different days.
abstract: This study was conducted to establish pig embryonic stem (ES)-like cell lines from nuclear transfer blastocysts. A green fluorescent protein (GFP)-expressing cell line was used as the source of donor cells injected into the enucleated oocytes. Blastocysts were collected at D5 (the fifth day), D7 (the seventh day) and D9 (the ninth day). Differential staining was used to assay the viability and development of blastocysts from the 3 days. The number of inner cell mass (ICM) cells increased from 1.83 ± 0.8 (D5) to 5.37 ± 1.2 (D7) to 7.56 ± 1.5 (D9). The expression profiles of embryonic stem (ES) cell factors (OCT4, SOX2, KLF4 and c-MYC) correlated best with the undifferentiated ES state and were identified by qPCR. The expression of the four factors was increased from D5 to D7, whereas the expression decreased from D7 to D9. We tried to isolate ES-like cells from these embryos. However, ES-like cells from the D7 blastocysts grew slowly and expressed alkaline phosphatase. The cells from the D9 blastocysts grew rapidly but did not express alkaline phosphatase. ES-like cells were not isolated from the D5 blastocysts. These results show that the cells from the D7 embryos are pluripotent but grow slowly. The cells from the D9 embryos grow rapidly but start to lose pluripotency.
authors: Jilin Province Animal Embryo Engineering Key Laboratory, College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China.
authors: Guangyun
authors: G
authors: Tan
authors: Linzhu
authors: L
authors: Ren
authors: Yongye
authors: Y
authors: Huang
authors: Xiaochun
authors: X
authors: Tang
authors: Yang
authors: Y
authors: Zhou
authors: Yan
authors: Y
authors: Zhou
authors: Dong
authors: D
authors: Li
authors: Hongxiao
authors: H
authors: Song
authors: Hongsheng
authors: H
authors: Ouyang
authors: Daxin
authors: D
authors: Pang
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D017348:Microscopy, Electron, Scanning Transmission; D013552:Swine / Q000196:embryology* / Q000378:metabolism