Porcine induced pluripotent stem cells require LIF and maintain their developmental potential in early stage of embryos

Porcine induced pluripotent stem (piPS) cell lines have been generated recently by using a cocktail of defined transcription factors, however, the features of authentic piPS cells have not been agreed upon and most of published iPS clones did not meet the stringent requirements of pluripotency. Here...

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Bibliographic Details
Published in:PloS one 2012-12, Vol.7 (12), p.e51778
Main Authors: Cheng, De, Guo, Yanjie, Li, Zhenzhen, Liu, Yajun, Gao, Xing, Gao, Yi, Cheng, Xiang, Hu, Junhe, Wang, Huayan
Format: Article
Language:English
Subjects:
Animals
Biology
Biotechnology
Blastocyst - cytology
Blastocyst - drug effects
Blastocysts
Cell Differentiation - drug effects
Cell lines
Cell Proliferation - drug effects
Cellular Reprogramming - drug effects
Chimera
Chimeras
Cloning
Cytology
DNA binding proteins
Embryo cells
Embryo, Mammalian - cytology
Embryo, Mammalian - drug effects
Embryonic stem cells
Embryos
Fibroblasts
Fibroblasts - cytology
Fibroblasts - drug effects
Fibroblasts - metabolism
Genetic engineering
Hogs
In vitro fertilization
Induced Pluripotent Stem Cells - cytology
Induced Pluripotent Stem Cells - drug effects
Induced Pluripotent Stem Cells - metabolism
Leukemia Inhibitory Factor - pharmacology
Mice
Morphology
Myc protein
Nuclear transfer
Nuclear Transfer Techniques
Penicillin
Pluripotency
Retroviridae
Stage-Specific Embryonic Antigens - metabolism
Stem cell transplantation
Stem cells
Sus scrofa - embryology
Transcription factors
Transduction, Genetic
Veterinary colleges
Veterinary medicine
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Summary:Porcine induced pluripotent stem (piPS) cell lines have been generated recently by using a cocktail of defined transcription factors, however, the features of authentic piPS cells have not been agreed upon and most of published iPS clones did not meet the stringent requirements of pluripotency. Here, we report the generation of piPS cells from fibroblasts using retrovirus carrying four mouse transcription factors (mOct4, mSox2, mKlf4 and mc-Myc, 4F). Multiple LIF-dependent piPS cell lines were generated and these cells showed the morphology similar to mouse embryonic stem cells and other pluripotent stem cells. In addition to the routine characterization, piPS cells were injected into porcine pre-compacted embryos to generate chimera embryos and nuclear transfer (NT) embryos. The results showed that piPS cells retain the ability to integrate into inner and outer layers of the blastocysts, and support the NT embryos development to blastocysts. The generations of chimera embryos and NT embryos derived from piPS clones are a practical means to determine the quality of iPS cells ex vivo.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0051778