Number of the records: 1  

Ex vivo tools for the clonal analysis of zebrafish hematopoiesis

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    SYSNO ASEP0473122
    Document TypeJ - Journal Article
    R&D Document TypeJournal Article
    Subsidiary JČlánek ve WOS
    TitleEx vivo tools for the clonal analysis of zebrafish hematopoiesis
    Author(s) Svoboda, Ondřej (UMG-J)
    Stachura, D.L. (US)
    Machoňová, Olga (UMG-J)
    Zon, L.I. (US)
    Traver, D. (US)
    Bartůněk, Petr (UMG-J) RID
    Number of authors6
    Source TitleNature Protocols. - : Nature Publishing Group - ISSN 1754-2189
    Roč. 11, č. 5 (2016), s. 1007-1020
    Number of pages14
    Languageeng - English
    CountryGB - United Kingdom
    Keywordsstem-cells ; in-vitro ; vertebrate hematopoiesis ; transgenic zebrafish ; progenitor cells ; danio-rerio ; fish ; thrombopoietin ; identification ; specification
    Subject RIVEB - Genetics ; Molecular Biology
    R&D ProjectsLO1419 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
    GA16-21024S GA ČR - Czech Science Foundation (CSF)
    Institutional supportUMG-J - RVO:68378050
    UT WOS000375154300003
    DOI10.1038/nprot.2016.053
    AnnotationThis protocol describes the ex vivo characterization of zebrafish hematopoietic progenitors. We show how to isolate zebrafish hematopoietic cells for cultivation and differentiation in colony assays in semi-solid media. We also describe procedures for the generation of recombinant zebrafish cytokines and for the isolation of carp serum, which are essential components of the medium required to grow zebrafish hematopoietic cells ex vivo. The outcome of these clonal assays can easily be evaluated using standard microscopy techniques after 3-10 d in culture. In addition, we describe how to isolate individual colonies for further imaging and gene expression profiling. In other vertebrate model organisms, ex vivo assays have been crucial for elucidating the relationships among hematopoietic stem cells (HSCs), progenitor cells and their mature progeny. The present protocol should facilitate such studies on cells derived from zebrafish.
    WorkplaceInstitute of Molecular Genetics
    ContactNikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217
    Year of Publishing2017
Number of the records: 1  

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