2018-02-162018-02-162011-06FREITAS, Érika Regina Leal de; SANCHES, Bruno Valente; GAMBARINI, Maria Lúcia; OLIVEIRA FILHO, Benedito Dias; GUILLO, Lidia Andreu. Embryonic stem-like cells derived from in vitro produced bovine blastocysts. Brazilian Archives of Biology and Technology, Curitiba, v. 54, n. 3, p. 495-502, May/June 2011.e- 1678-43241516-8913http://repositorio.bc.ufg.br/handle/ri/13650The aim of this work was to study the derivation of bovine embryonic stem-like (ES-like) cells from the inner cell mass (ICM) of in vitro produced blastocysts. The ICMs were mechanically isolated and six out of seventeen (35%) ICMs could attach to a monolayer of murine embryonic fibroblasts (MEF).Ten days after, primary outgrowths were mechanically dissected into several small clumps and transferred to a new MEF layer. Cells were further propagated and passaged by physical dissociation over a 60 days period. The pluripotency of the bovine ES-like cells was confirmed by RT-PCR of Oct-4 and STAT-3 gene markers. The colonies were weakly stained for alkaline phosphatase and the mesoderm and endoderm differentiation gene markers such as GATA-4 and Flk-1, respectively, were not expressed. Embryoid bodies were spontaneously formed at the seventh passage. Results showed that bovine ES-like cells could be obtained and passaged by mechanical procedures from the fresh in vitro produced blastocysts.engAcesso AbertoBovine blastocystsICM isolationEmbryonic stem cellsGene markersArtigo10.1590/S1516-89132011000300009