Back to Search Start Over

Derivation of Porcine Embryonic Stem-Like Cells from In Vitro-Produced Blastocyst-Stage Embryos

Derivation of Porcine Embryonic Stem-Like Cells from In Vitro-Produced Blastocyst-Stage Embryos

Authors :
Qing-Yuan Sun
Lihua Zhao
Yuan Chen
Ning Yang
Yifan Dai
Yong Jin
Yan-Ru Li
Haibin Jiang
Rongfeng Li
Xiaowei Nie
Manling Zhang
Zhaoqiang Wu
Na Ta
Daorong Hou
Source :
Scientific Reports
Publication Year :
2016
Publisher :
Springer Science and Business Media LLC, 2016.

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.

Details

ISSN :
20452322
Volume :
6
Database :
OpenAIRE
Journal :
Scientific Reports
Accession number :
edsair.doi.dedup.....2caac7f179cadbd56aafde248e579f6c
Full Text :
https://doi.org/10.1038/srep25838