Embryonic stem-like cells derived from in vitro produced bovine blastocysts

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). T...

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Bibliographic Details
Main Authors: Erika Regina Leal de Freitas, Bruno Valente Sanches, Maria Lúcia Gambarini, Benedito Dias Oliveira Filho, Lidia Andreu Guillo
Format: Article
Language:English
Published: Instituto de Tecnologia do Paraná (Tecpar) 2011-06-01
Series:Brazilian Archives of Biology and Technology
Subjects:
Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132011000300009
Description
Summary: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.
ISSN:1516-8913
1678-4324