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Isolation and Culture of Bovine Embryonic Stem Cells

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Epiblast Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1074))

Abstract

Isolation and culture of primary embryonic stem (ES) cell colonies are the first critical step towards establishment of stable ES cell lines. Here we introduce a novel method designated as “Separate and Seed” that contributes remarkably to efficient derivation of bovine primary ES cell colonies from blastocysts. The bovine ES cell colonies can self-renew to passage 10 with the growth factors bFGF and BIO. The bovine ES cells exhibit morphology typical of ES cells and express pluripotent molecular markers including Oct4, Nanog, SSEA1, SSEA4, and alkaline phosphatase (AP). These pluripotent markers may be used for the characterization of authentic bovine ES cell lines. Although continued efforts are required for improving long-term culture of bovine ES cells, this novel “Separate and Seed” method plus the growth factors bFGF and BIO provides an initial effective step that may eventually lead to the derivation of authentic bovine ES cells.

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Acknowledgements

This study was supported by Ministry of Science and Technology China (863 plan 2010AA10A103), the National Basic Research Program of China (2009CB941000 and 2011CBA01002) and China National Natural Science Foundation.

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Authors and Affiliations

  1. The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, China

    Shanbo Cao

  2. State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, China

    Fang Wang

  3. Department of Cell Biology and Genetics, College of Life Sciences, Nankai University, Tianjin, China

    Fang Wang

  4. College of Life Sciences, Nankai University, Tianjin, China

    Lin Liu

Authors
  1. Shanbo Cao
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  2. Fang Wang
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  3. Lin Liu
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Editor information

Editors and Affiliations

  1. School of Biosciences Rm 223 South Lab, University of Nottingham, Loughborough, United Kingdom

    Ramiro Alberio

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Cao, S., Wang, F., Liu, L. (2013). Isolation and Culture of Bovine Embryonic Stem Cells. In: Alberio, R. (eds) Epiblast Stem Cells. Methods in Molecular Biology, vol 1074. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-628-3_9

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  • DOI: https://doi.org/10.1007/978-1-62703-628-3_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-627-6

  • Online ISBN: 978-1-62703-628-3

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