title: Trichostatin A-mediated epigenetic transformation of adult bone marrow-derived mesenchymal stem cells biases the in vitro developmental capability, quality, and pluripotency extent of porcine cloned embryos.
abstract: The current research was conducted to explore the in vitro developmental outcome and cytological/molecular quality of porcine nuclear-transferred (NT) embryos reconstituted with adult bone marrow-derived mesenchymal stem cells (ABM-MSCs) that were epigenetically transformed by treatment with nonspecific inhibitor of histone deacetylases, known as trichostatin A (TSA). The cytological quality of cloned blastocysts was assessed by estimation of the total cells number (TCN) and apoptotic index. Their molecular quality was evaluated by real-time PCR-mediated quantification of gene transcripts for pluripotency- and multipotent stemness-related markers (Oct4, Nanog, and Nestin). The morula and blastocyst formation rates of NT embryos derived from ABM-MSCs undergoing TSA treatment were significantly higher than in the TSA-unexposed group. Moreover, the NT blastocysts generated using TSA-treated ABM-MSCs exhibited significantly higher TCN and increased pluripotency extent measured with relative abundance of Oct4 and Nanog mRNAs as compared to the TSA-untreated group. Altogether, the improvements in morula/blastocyst yields and quality of cloned pig embryos seem to arise from enhanced abilities for promotion of correct epigenetic reprogramming of TSA-exposed ABM-MSC nuclei in a cytoplasm of reconstructed oocytes. To our knowledge, we are the first to report the successful production of mammalian high-quality NT blastocysts using TSA-dependent epigenomic modulation of ABM-MSCs.
authors: Department of Biotechnology of Animal Reproduction, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice n. Kraków, Poland.
authors: Marcin
authors: M
authors: Samiec
authors: Department of Biotechnology of Animal Reproduction, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice n. Kraków, Poland.
authors: Jolanta
authors: J
authors: Opiela
authors: Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Dojazd 11 Street, 60-632 Poznań, Poland ; Institute of Human Genetics, Polish Academy of Sciences, Strzeszyńska 32 Street, 60-479 Poznań, Poland.
authors: Daniel
authors: D
authors: Lipiński
authors: Department of Biotechnology of Animal Reproduction, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice n. Kraków, Poland.
authors: Joanna
authors: 0000-0001-9068-4300
authors: J
authors: Romanek
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D019976:Cloning, Organism; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005260:Female; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D013552:Swine
chemical_list: D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; C012589:trichostatin A