PUBLICATION
Zebrafish kidney marrow contains ABCG2-dependent side population cells exhibiting hematopoietic stem cell properties
- Authors
- Tsinkalovsky, O., Vik-Mo, A.O., Ferreira, S., Laerum, O.D., and Fjose, A.
- ID
- ZDB-PUB-070212-41
- Date
- 2007
- Source
- Differentiation; research in biological diversity 75(3): 175-183 (Journal)
- Registered Authors
- Ferreira, Sara, Fjose, Anders
- Keywords
- none
- MeSH Terms
-
- ATP-Binding Cassette Transporters/antagonists & inhibitors
- ATP-Binding Cassette Transporters/metabolism*
- Animals
- Benzimidazoles/metabolism
- Cell Differentiation
- Flow Cytometry
- Hematopoietic Stem Cells/cytology*
- Hematopoietic Stem Cells/drug effects
- Hematopoietic Stem Cells/metabolism
- Indoles/pharmacology
- Kidney/cytology*
- Kidney/drug effects
- Kidney/metabolism
- Phenotype
- Reserpine/pharmacology
- Zebrafish/metabolism*
- Zebrafish Proteins/antagonists & inhibitors
- Zebrafish Proteins/metabolism*
- PubMed
- 17288542 Full text @ Differentiation
Citation
Tsinkalovsky, O., Vik-Mo, A.O., Ferreira, S., Laerum, O.D., and Fjose, A. (2007) Zebrafish kidney marrow contains ABCG2-dependent side population cells exhibiting hematopoietic stem cell properties. Differentiation; research in biological diversity. 75(3):175-183.
Abstract
Zebrafish (Danio rerio) has emerged as a powerful genetic model for the study of vertebrate hematopoiesis. However, methods for detection and isolation of hematopoietic stem cells (HSCs) have not yet been reported. In mammals, the combination of Hoechst 33342 staining with flow cytometry can be used for separation of a bone marrow side population (SP), which is highly enriched for HSCs. We applied a similar procedure to hematopoietic kidney marrow cells from adult zebrafish, and identified a segregated cohort of SP cells, which demonstrate a set of features typical of stem cells. SP cells show extremely low scatter characteristics, and are small in size with a minimum of cytoplasm. Treatment of zebrafish kidney marrow cells with reserpine or fumitremorgin C, which inhibit the ABCG2 transporter responsible for Hoechst 33342 efflux, caused a clear reduction in the number of SP cells. Consistent with the quiescent state of HSCs, the SP cells are strongly resistant to the myelosuppressive agent 5-fluorouracil. In addition, SP cells specifically demonstrate higher expression of genes known to be markers of HSCs of mammals. Hence, our results show that the SP phenotype is conserved between mammals and teleosts, and the properties of the zebrafish SP cells indicate a significant enrichment for HSCs. These rapid flow cytometric methods for purification of HSCs from zebrafish may greatly facilitate genetic analysis of stem cells using the advantages of this vertebrate model.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping