title: Development and gene expression of porcine cloned embryos derived from bone marrow stem cells with overexpressing Oct4 and Sox2.
abstract: The present study compared the potential of porcine bone marrow mesenchymal stem cells (pBMSCs) at different passages as nuclear transfer (NT) donors and the developmental efficiency of NT embryos from donor cells transfected with/without Oct4 and Sox2. Early-passage pBMSCs showed higher proliferation and expression of Oct4 and Sox2 and differentiation potential into mesenchymal lineages than middle- and late-passage pBMSCs. Cleavage rate did not differ among pBMSCs at different passages, but NT embryos with early-passage pBMSCs and middle-passage pBMSCs transfected with Oct4 (Oct4-pBMSCs) had significantly (p<0.05) higher blastocyst development than those with middle-passage pBMSCs. The incidence of apoptotic bodies in NT blastocysts from late-passage pBMSCs and Sox2-transfected middle-passage pBMSCs (Sox2-pBMSCs) was significantly (p<0.05) higher than others. The transcriptional levels of Oct4, Sox2, Nanog, Cdx2, Dnmt3a, and Igf2r genes were significantly (p<0.05) higher in Oct4- and Sox2-pBMSCs NT embryos. Middle-passage pBMSCs NT embryos revealed lower transcriptional levels of Bcl2 than others, except Sox2-pBMSCs NT embryos. The transcriptional level of Bax increased gradually in NT embryos derived from pBMSCs following extended passages and was significantly (p<0.05) higher in Sox2-pBMSCs NT embryos. Our results demonstrated that early-passage pBMSCs are more potent in expressing transcription factors and displayed higher differentiation ability, and middle-passage pBMSCs transfected with Oct4 improved the developmental efficiency of NT embryos, suggesting that high Oct4 expression cells are more efficient as NT donors.
authors.affiliation: 1 Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University , Jinju, 660-701, Republic of Korea.
authors.forename: Jeong-Hyeon
authors.initials: JH
authors.lastname: Lee
authors.forename: Won-Jae
authors.initials: WJ
authors.lastname: Lee
authors.forename: Ryoung-Hoon
authors.initials: RH
authors.lastname: Jeon
authors.forename: Yeon-Mi
authors.initials: YM
authors.lastname: Lee
authors.forename: Si-Jung
authors.initials: SJ
authors.lastname: Jang
authors.forename: Sung-Lim
authors.initials: SL
authors.lastname: Lee
authors.forename: Byung-Geun
authors.initials: BG
authors.lastname: Jeon
authors.forename: Sun-A
authors.initials: SA
authors.lastname: Ock
authors.forename: W Allen
authors.initials: WA
authors.lastname: King
authors.forename: Gyu-Jin
authors.initials: GJ
authors.lastname: Rho
mesh_terms: D000818:Animals; D017209:Apoptosis; D002454:Cell Differentiation; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods*; D046149:Embryo Culture Techniques / Q000662:veterinary; D047108:Embryonic Development; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism*; D013552:Swine; D014162:Transfection
chemical_list: D018398:Homeodomain Proteins; D050814:Octamer Transcription Factor-3; D055748:SOXB1 Transcription Factors