title: Zebrafish embryonic stromal trunk (ZEST) cells support hematopoietic stem and progenitor cell (HSPC) proliferation, survival, and differentiation.
abstract: Forward genetic screens in zebrafish have been used to identify genes essential for the generation of primitive blood and the emergence of hematopoietic stem cells (HSCs), but have not elucidated the genes essential for hematopoietic stem and progenitor cell (HSPC) proliferation and differentiation because of the lack of methodologies to functionally assess these processes. We previously described techniques used to test the developmental potential of HSPCs by culturing them on zebrafish kidney stromal (ZKS) cells, derived from the main site of hematopoiesis in the adult teleost. Here we describe an additional primary stromal cell line we refer to as zebrafish embryonic stromal trunk (ZEST) cells, derived from tissue surrounding the embryonic dorsal aorta, the site of HSC emergence in developing fish. ZEST cells encouraged HSPC differentiation toward the myeloid, lymphoid, and erythroid pathways when assessed by morphologic and quantitative reverse transcription polymerase chain reaction analyses. Additionally, ZEST cells significantly expanded the number of cultured HSPCs in vitro, indicating that these stromal cells are supportive of both HSPC proliferation and multilineage differentiation. Examination of ZEST cells indicates that they express numerous cytokines and Notch ligands and possess endothelial characteristics. Further characterization of ZEST cells should prove to be invaluable in understanding the complex signaling cascades instigated by the embryonic hematopoietic niche required to expand and differentiate HSPCs. Elucidating these processes and identifying possibilities for the modulation of these molecular pathways should allow the in vitro expansion of HSPCs for a multitude of therapeutic uses.
authors.affiliation: Department of Cellular and Molecular Medicine, University of California at San Diego School of Medicine, La Jolla, California, USA.
authors.forename: Clyde
authors.initials: C
authors.lastname: Campbell
authors.affiliation: Department of Cellular and Molecular Medicine, University of California at San Diego School of Medicine, La Jolla, California, USA.
authors.forename: Tammy
authors.initials: T
authors.lastname: Su
authors.affiliation: Department of Cellular and Molecular Medicine, University of California at San Diego School of Medicine, La Jolla, California, USA.
authors.forename: Ryan P
authors.initials: RP
authors.lastname: Lau
authors.affiliation: Department of Biological Sciences, California State University, Chico, California, USA.
authors.forename: Arpit
authors.initials: A
authors.lastname: Shah
authors.affiliation: Department of Biological Sciences, California State University, Chico, California, USA.
authors.forename: Payton C
authors.initials: PC
authors.lastname: Laurie
authors.affiliation: Department of Biological Sciences, California State University, Chico, California, USA.
authors.forename: Brenda
authors.initials: B
authors.lastname: Avalos
authors.affiliation: Department of Biological Sciences, California State University, Chico, California, USA.
authors.forename: Julian
authors.initials: J
authors.lastname: Aggio
authors.affiliation: Department of Biological Sciences, California State University, Chico, California, USA.
authors.forename: Elena
authors.initials: E
authors.lastname: Harris
authors.affiliation: Department of Cellular and Molecular Medicine, University of California at San Diego School of Medicine, La Jolla, California, USA.
authors.forename: David
authors.initials: D
authors.lastname: Traver
authors.affiliation: Department of Biological Sciences, California State University, Chico, California, USA. Electronic address: dstachura@csuchico.edu.
authors.forename: David L
authors.initials: DL
authors.lastname: Stachura
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology*; D002470:Cell Survival / Q000502:physiology; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism*; D015027:Zebrafish / Q000378:metabolism*