Embryonic stem-like cells derived from in vitro produced bovine blastocysts
AUTOR(ES)
Freitas, Erika Regina Leal de, Sanches, Bruno Valente, Gambarini, Maria Lúcia, Oliveira Filho, Benedito Dias, Guillo, Lidia Andreu
FONTE
Brazilian Archives of Biology and Technology
DATA DE PUBLICAÇÃO
2011-06
RESUMO
The 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.
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