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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
- 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.
- Subjects :
- Pluripotent Stem Cells
0301 basic medicine
Homeobox protein NANOG
KOSR
Transcription, Genetic
Rex1
Sus scrofa
Fertilization in Vitro
Embryoid body
Biology
Article
Colony-Forming Units Assay
03 medical and health sciences
0302 clinical medicine
SOX2
medicine
Animals
Cluster Analysis
Blastocyst
Cell Shape
Embryoid Bodies
Embryonic Stem Cells
reproductive and urinary physiology
030219 obstetrics & reproductive medicine
Multidisciplinary
Gene Expression Profiling
Teratoma
Cell Differentiation
Molecular biology
Embryonic stem cell
Cell biology
030104 developmental biology
medicine.anatomical_structure
Gene Expression Regulation
embryonic structures
Stem cell
Biomarkers
Subjects
Details
- ISSN :
- 20452322
- Volume :
- 6
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....2caac7f179cadbd56aafde248e579f6c
- Full Text :
- https://doi.org/10.1038/srep25838