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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.

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