PUBLICATION

Enumerating Hematopoietic Stem and Progenitor Cells in Zebrafish Embryos

Authors
Esain, V., Cortes, M., North, T.E.
ID
ZDB-PUB-160729-17
Date
2016
Source
Methods in molecular biology (Clifton, N.J.)   1451: 191-206 (Chapter)
Registered Authors
North, Trista
Keywords
CFU-C assay, Flow cytometry, Hematopoietic stem cells, Live imaging, Zebrafish embryo
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Cell Differentiation/genetics
  • Cell Differentiation/physiology
  • Embryo, Nonmammalian/cytology
  • Embryo, Nonmammalian/metabolism
  • Flow Cytometry
  • Hematopoiesis/genetics
  • Hematopoiesis/physiology*
  • Hematopoietic Stem Cells/cytology*
  • Hematopoietic Stem Cells/metabolism*
  • Signal Transduction/genetics
  • Signal Transduction/physiology
  • Zebrafish/embryology*
  • Zebrafish/metabolism*
PubMed
27464809 Full text @ Meth. Mol. Biol.
Abstract
Over the past 20 years, zebrafish have proven to be a valuable model to dissect the signaling pathways involved in hematopoiesis, including Hematopoietic Stem and Progenitor Cell (HSPC) formation and homeostasis. Despite tremendous efforts to generate the tools necessary to characterize HSPCs in vitro and in vivo the zebrafish community still lacks standardized methods to quantify HSPCs across laboratories. Here, we describe three methods used routinely in our lab, and in others, to reliably enumerate HSPCs in zebrafish embryos: large-scale live imaging of transgenic reporter lines, Fluorescence-Activated Cell Sorting (FACS), and in vitro cell culture. While live imaging and FACS analysis allows enumeration of total or site-specific HSPCs, the cell culture assay provides the unique opportunity to test the functional potential of isolated HSPCs, similar to those employed in mammals.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping