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

PLoS One. 2012;7(12):e51778. doi: 10.1371/journal.pone.0051778. Epub 2012 Dec 14.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blastocyst / cytology
  • Blastocyst / drug effects
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Cellular Reprogramming / drug effects
  • Chimera
  • Embryo, Mammalian / cytology*
  • Embryo, Mammalian / drug effects
  • Fibroblasts / cytology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism
  • Leukemia Inhibitory Factor / pharmacology*
  • Mice
  • Nuclear Transfer Techniques
  • Retroviridae
  • Stage-Specific Embryonic Antigens / metabolism
  • Sus scrofa / embryology*
  • Transduction, Genetic

Substances

  • Leukemia Inhibitory Factor
  • Stage-Specific Embryonic Antigens
  • stage-specific embryonic antigen-4

Grant support

This work was supported by The National Basic Research Program of China (2009CB941002). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.