Papers

Peer-reviewed
10 2011

Analysis of Marker Expression in Porcine Cell Lines Derived from Blastocysts Produced In Vitro and In Vivo

JOURNAL OF REPRODUCTION AND DEVELOPMENT
  • Irena Vackova
  • ,
  • Zora Novakova
  • ,
  • Vladimir Krylov
  • ,
  • Konosuke Okada
  • ,
  • Tomas Kott
  • ,
  • Helena Fulka
  • ,
  • Jan Motlik

Volume
57
Number
5
First page
594
Last page
603
Language
English
Publishing type
Research paper (scientific journal)
DOI
10.1262/jrd.10-184H
Publisher
SOCIETY REPRODUCTION & DEVELOPMENT-SRD

The present study was designed to extensively characterize cell lines derived from porcine blastocysts by several methodical approaches, including morphological observation, cytogenetic analysis, estimation of alkaline phosphatase activity and detection of specific marker expression at the mRNA/protein level. A comparison was made between the properties of cell lines isolated from in vivo- and in vitro-obtained blastocysts. Our results showed that 57.1% of the in vivo-obtained blastocysts attached to the feeder layer and that 33.3% of them started to grow in a monolayer. The percentage of attached in vitro-produced blastocysts was lower (24.6%), and only 6.9% of them started to grow. Outgrowths from the in vitro-produced blastocysts formed mainly trophectoderm or epithelial-like monolayer, whereas the in vivo-obtained blastocysts formed heterogeneous outgrowths that also contained cells with embryonic stem (ES)-like morphology. Detailed analyses showed that the primary outgrowths with ES-like morphology expressed the pluripotency markers OCT-4 and NANOG and revealed intensive alkaline phosphatase staining, while they did not express markers of differentiation. The majority of passaged cells, including those with ES-like morphology, lacked OCT-4 protein and revealed expression of specific differentiation markers (CYTOKERATIN 18, LAMINS A/C, TRANSFERRIN, a-FETOPROTEIN and GATA-4), although they still expressed NANOG and exhibited weak alkaline phosphatase activity. Moreover, these cells spontaneously differentiated into neural, fibroblast or epithelial-like cells, even in the presence of leukaemia inhibitory factor. Our results show that complex analysis of markers of pluripotency as well as differentiation markers is necessary for proper interpretation of data in porcine embryonic stem cell studies.

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Link information
DOI
https://doi.org/10.1262/jrd.10-184H
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000296597100006&DestApp=WOS_CPL

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