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Ex vivo tools for the clonal analysis of zebrafish hematopoiesis

O. Svoboda, DL. Stachura, O. Machonova, LI. Zon, D. Traver, P. Bartunek,

. 2016 ; 11 (5) : 1007-20. [pub] 20160428

Jazyk angličtina Země Anglie, Velká Británie

Typ dokumentu časopisecké články, Research Support, N.I.H., Extramural, práce podpořená grantem

Perzistentní odkaz   https://www.medvik.cz/link/bmc16027538

This 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.

Citace poskytuje Crossref.org

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$a Stachura, David L $u Department of Biological Sciences, California State University Chico, Chico, California, USA.
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$a Zon, Leonard I $u Stem Cell Program, Dana-Farber/Boston Children's, Boston, Massachusetts, USA. Division of Hematology/Oncology, Dana-Farber/Boston Children's, Boston, Massachusetts, USA. Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical School, Boston, Massachusetts, USA.
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$a Traver, David $u Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California, USA.
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