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

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概要

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• 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, α-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.

著者

• FULKA Helena

  Institute of Animal Science

• VACKOVA Irena

  Institute of Animal Science

• NOVAKOVA Zora

  Institute of Animal Science

• KRYLOV Vladimir

  Institute of Animal Science

• OKADA Konosuke

  Institute of Animal Science

• KOTT Tomas

  Institute of Animal Science

• MOTLIK Jan

  Center for Cell Therapy and Tissue Repair

関連論文

• Epigenetic Characteristics of Paternal Chromatin in Interspecies Zygotes
• 単離されたマウス卵核胞の凍結保存
• Analysis of Marker Expression in Porcine Cell Lines Derived from Blastocysts Produced In Vitro and In Vivo
• Epigenetic Characteristics of Paternal Chromatin in Interspecies Zygotes
• How to Repair the Oocyte and Zygote?
• Putative Embryonic Stem Cell Lines from Pig Embryos
• Analysis of Marker Expression in Porcine Cell Lines Derived from Blastocysts Produced In Vitro and in Vivo
• Production of Giant Mouse Oocyte Nucleoli and Assessment of Their Protein Content
• Cryopreservation of Isolated Mouse Germinal Vesicles

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