Induced pluripotent stem cells from goat fibroblasts

Mol Reprod Dev. 2013 Dec;80(12):1009-17. doi: 10.1002/mrd.22266. Epub 2013 Nov 27.

Abstract

Embryonic stem cells (ESCs) are a powerful model for genetic engineering, studying developmental biology, and modeling disease. To date, ESCs have been established from the mouse (Evans and Kaufman, 1981, Nature 292:154-156), non-human primates (Thomson et al., , Proc Nat Acad Sci USA 92:7844-7848), humans (Thomson et al., 1998, Science 282:1145-1147), and rats (Buehr et al., , Cell 135:1287-1298); however, the derivation of ESCs from domesticated ungulates such as goats, sheep, cattle, and pigs have not been successful. Alternatively, induced pluripotent stem cells (iPSCs) can be generated by reprogramming somatic cells with several combinations of genes encoding transcription factors (OCT3/4, SOX2, KLF4, cMYC, LIN28, and NANOG). To date, iPSCs have been isolated from various species, but only limited information is available regarding goat iPSCs (Ren et al., 2011, Cell Res 21:849-853). The objectives of this study were to generate goat iPSCs from fetal goat primary ear fibroblasts using lentiviral transduction of four human transcription factors: OCT4, SOX2, KLF4, and cMYC. The goat iPSCs were successfully generated by co-culture with mitomycin C-treated mouse embryonic fibroblasts using medium supplemented with knockout serum replacement and human basic fibroblast growth factor. The goat iPSCs colonies are flat, compact, and closely resemble human iPSCs. They have a normal karyotype; stain positive for alkaline phosphatase, OCT4, and NANOG; express endogenous pluripotency genes (OCT4, SOX2, cMYC, and NANOG); and can spontaneously differentiate into three germ layers in vitro and in vivo.

MeSH terms

  • Alkaline Phosphatase / biosynthesis
  • Animals
  • Cell Differentiation
  • Cells, Cultured
  • Cellular Reprogramming / genetics*
  • Coculture Techniques
  • Fibroblast Growth Factor 2 / pharmacology
  • Fibroblasts / cytology
  • Goats
  • HEK293 Cells
  • Homeodomain Proteins / biosynthesis
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Karyotype
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors / genetics
  • Mice
  • Mice, Inbred C57BL
  • Mitomycin / pharmacology
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3 / biosynthesis
  • Octamer Transcription Factor-3 / genetics
  • Proto-Oncogene Proteins c-myc / biosynthesis
  • Proto-Oncogene Proteins c-myc / genetics
  • SOXB1 Transcription Factors / biosynthesis
  • SOXB1 Transcription Factors / genetics

Substances

  • Homeodomain Proteins
  • KLF4 protein, human
  • Klf4 protein, mouse
  • Klf4 protein, rat
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors
  • MYC protein, human
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • Proto-Oncogene Proteins c-myc
  • SOX2 protein, human
  • SOXB1 Transcription Factors
  • Fibroblast Growth Factor 2
  • Mitomycin
  • Alkaline Phosphatase