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). 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.

Saved in:
Bibliographic Details
Main Authors: Freitas,Erika Regina Leal de, Sanches,Bruno Valente, Gambarini,Maria Lúcia, Oliveira Filho,Benedito Dias, Guillo,Lidia Andreu
Format: Digital revista
Language:English
Published: Instituto de Tecnologia do Paraná - Tecpar 2011
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132011000300009
Tags: Add Tag
No Tags, Be the first to tag this record!