title: Constitutive expression of the embryonic stem cell marker OCT4 in bovine somatic donor cells influences blastocysts rate and quality after nucleus transfer.
abstract: Nuclear transfer (NT) is associated with epigenetic reprogramming of donor cells. Expression of certain genes in these cells might facilitate their expression in the NT embryo. This research was aimed to investigate the effect of constitutive expression of OCT4 in bovine somatic cells used for NT on the developmental potential of derived cloned embryos as well as in the expression of pluripotency markers in the Day-7 resulting embryos. Cloned blastocysts were generated from five cell lines that expressed OCT4. Pools of blastocysts were screened to detect OCT4, SOX2, and NANOG by qPCR. In vitro-fertilized time-matched blastocysts were used as controls. The development potential was assessed on the basis of blastocysts rate; grading and total cell counts at Day 7. OCT4 expression in the cell lines positively correlates with blastocysts rate (r = 0.92; p = 0.02), number of grade I blastocysts (r = 0.96; p = 0.01), and total cell number (r = 0.98; p = 0.002). The high expression of OCT4 in the cell line did not improve the final outcome of cloning. Somatic expression of OCT4 lead to increased expression of OCT4 and SOX2 in cloned grade I blastocysts; however, there was a bigger variability in OCT4 and SOX2 (p = 0.03; p = 0.02) expression in the embryos generated from cells expressing highest levels of OCT4. Probably the higher variability in OCT4 expression in cloned embryos is due to incorrect reprogramming and incapability of the oocyte to correct for higher OCT4 levels. For that reason, we concluded that OCT4 expression in somatic cells is not a good prognosis marker for selecting cell lines.
authors.affiliation: Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepcion, Avenida Vicente Mendez 595, Chillán, Chile, llrodriguez@udec.cl.
authors.forename: Lleretny
authors.initials: L
authors.lastname: Rodríguez-Alvarez
authors.forename: Jose
authors.initials: J
authors.lastname: Manriquez
authors.forename: Alejandra
authors.initials: A
authors.lastname: Velasquez
authors.forename: Fidel Ovidio
authors.initials: FO
authors.lastname: Castro
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002417:Cattle; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D004622:Embryo, Mammalian; D047108:Embryonic Development; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental; D053652:Nuclear Transfer Techniques; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis*; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism
chemical_list: D050814:Octamer Transcription Factor-3