title: Exposure of Somatic Cells to Cytoplasm Extracts of Porcine Oocytes Induces Stem Cell-Like Colony Formation and Alters Expression of Pluripotency and Chromatin-Modifying Genes.
abstract: Cell permeabilization followed by exposure to cytoplasmic extracts of oocytes has been proposed as an alternative to transduction of transcription factors for inducing pluripotency in cultured somatic cells. The main goal in this study was to investigate the effect of treating porcine fibroblast cells with cytoplasmic extracts of GV-stage oocyte (OEx) followed by inhibition of histone deacetylases with Scriptaid (Scrip) on the formation of stem cell-like colonies and expression of genes encoding pluripotency and chromatin-modifying enzymes. Stem cell-like colonies start developing ∼2 weeks after treatment in cells exposed to OEx or OEx + Scrip. The number of cell colonies at the first day of appearance and 48 hours later was also similar between OEx and OEx + Scrip treatments. Transcripts for Nanog, Rex1, and c-Myc genes were detected in most cell samples that were analyzed on different days after OEx treatment. However, Sox2 transcripts were not detected and only a small proportion of samples had detectable levels of Oct4 mRNA after OEx treatment. A similar pattern of transcripts for pluripotency genes was observed in cells treated with OEx alone or OEx + Scrip. Transcript levels for Dnmt1 and Ezh2 were reduced at Day 3 after treatment in cells exposed to OEx. These findings revealed that: (a) exposure to OEx can induce a partial reprogramming of fibroblast cells toward pluripotency, characterized by colony formation and activation of pluripotency genes; and (b) inhibition of histone deacetylases does not improve the reprogramming effect of OEx treatment.
authors.affiliation: 1 Laboratory of Biotechnology and Animal Reproduction-BioRep, Federal University of Santa Maria (UFSM) , Santa Maria, Brazil .
authors.forename: Werner Giehl
authors.initials: WG
authors.lastname: Glanzner
authors.affiliation: 2 Postgraduate Program in Biotechnology, Laboratory of Molecular Embryology and Transgenesis, Technology Development Center, Federal University of Pelotas (UFPEL) , Pelotas, Brazil .
authors.forename: Eliza R
authors.initials: ER
authors.lastname: Komninou
authors.affiliation: 3 Department of Animal Science, McGill University , Ste-Anne-De-Bellevue, Canada .
authors.forename: Ashwini
authors.initials: A
authors.lastname: Mahendran
authors.affiliation: 1 Laboratory of Biotechnology and Animal Reproduction-BioRep, Federal University of Santa Maria (UFSM) , Santa Maria, Brazil .
authors.forename: Vitor B
authors.initials: VB
authors.lastname: Rissi
authors.affiliation: 3 Department of Animal Science, McGill University , Ste-Anne-De-Bellevue, Canada .
authors.forename: Karina
authors.initials: K
authors.lastname: Gutierrez
authors.affiliation: 3 Department of Animal Science, McGill University , Ste-Anne-De-Bellevue, Canada .
authors.forename: Rodrigo C
authors.initials: RC
authors.lastname: Bohrer
authors.affiliation: 2 Postgraduate Program in Biotechnology, Laboratory of Molecular Embryology and Transgenesis, Technology Development Center, Federal University of Pelotas (UFPEL) , Pelotas, Brazil .
authors.forename: Tiago
authors.initials: T
authors.lastname: Collares
authors.affiliation: 1 Laboratory of Biotechnology and Animal Reproduction-BioRep, Federal University of Santa Maria (UFSM) , Santa Maria, Brazil .
authors.forename: Paulo B D
authors.initials: PB
authors.lastname: Gonçalves
authors.affiliation: 3 Department of Animal Science, McGill University , Ste-Anne-De-Bellevue, Canada .
authors.forename: Vilceu
authors.initials: V
authors.lastname: Bordignon
mesh_terms: D000818:Animals; D002457:Cell Extracts / Q000494:pharmacology*; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D042002:Chromatin Assembly and Disassembly; D005347:Fibroblasts / Q000166:cytology; D020869:Gene Expression Profiling; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006898:Hydroxylamines / Q000494:pharmacology*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000737:chemistry*; D011804:Quinolines / Q000494:pharmacology*; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D013552:Swine
chemical_list: D002457:Cell Extracts; D056572:Histone Deacetylase Inhibitors; D006898:Hydroxylamines; D050814:Octamer Transcription Factor-3; D011804:Quinolines; D055748:SOXB1 Transcription Factors; C410733:scriptaid