Zebrafish stromal cells have endothelial properties and support hematopoietic cells

Abstract

Objective The goal of this study was to determine if we could establish a mesenchymal stromal line from zebrafish that would support hematopoietic cells. Such a co-culture system would be a great benefit to study the hematopoietic cell-stromal cell interaction in both the in vitro and in vivo environments. Methods Zebrafish stromal cells, ZStrC, were isolated from the “mesenchymal” tissue of the caudal tail and expanded in a specialized growth media. ZStrC were evaluated for phenotype, gene expression, and the ability to maintain zebrafish marrow cells in co-culture experiments. Results ZStrC showed mesenchymal and endothelial gene expression. Although ZStrC lacked the ability to differentiate into classic MSC lineages (osteocytes, adipocytes, chondrocytes), they did have the capacity for endotube formation on matrigel and LDL-uptake. ZStrC supported marrow cells for greater than 2 weeks in vitro. Importantly, the marrow cells were shown to retain homing ability in adoptive transfer experiments. ZStrC also were shown to improve hematopoietic recovery after sub-lethal irradiation after adoptive transfer. Conclusion As the zebrafish model grows in popularity and importance in the study of hematopoiesis, new tools to aid in our understanding of the hematopoietic cell-stromal cell interaction are required. ZStrC represent an additional tool in the study of hematopoiesis and will be useful to understand the factors that mediate the stromal cell-hematopoietic cell interaction that are important in hematopoietic maintenance.

Migration of cardiomyocytes is essential for heart regeneration in zebrafish

“…This contrasts to mammalian myocardial infarction models (Takahashi, 2010), according to which Cxcr4 is expressed in cell types involved in neovascularization, such as bone-marrow-derived mesenchymal stromal cells (Honczarenko et al, 2006;Yamaguchi et al, 2003) and endothelial progenitor cells (Yamaguchi et al, 2003). Although zebrafish stromal cells can exhibit endothelial-like properties (Lund et al, 2012), neo-vascularization seems to be unaffected in CXCR4-antagonist treated fish (Fig. 8).…”

“…S7). Moreover, it is yet to be determined whether mesenchymal stromal cells in zebrafish (Lund et al, 2012) can contribute to CMs. Nonetheless, our data highlight the requirement of Cxcr4 function in CMs during heart regeneration in zebrafish.…”

sdf1 Expression Reveals a Source of Perivascular-Derived Mesenchymal Stem Cells in Zebrafish

“…hMSCs are limited in the number of cell divisions they can undergo with a limit of around 50 population doublings . Whereas in zMSC (and other zebrafish stromal cell lines) there is no declination in doubling time nor change in cellular morphology even after 1–2 years in culture reflecting over 100 population doubling times . This phenomenon is most likely related to the constitutive activation of the telomerase gene that we and others have described in zebrafish cells .…”

“…A criterion for MSC is plastic adherence after isolation from the tissue source being tested (whether bone marrow, fat, or umbilical cord blood in origin), and subsequent tissue culture outgrowth . We and others have previously developed successful culture systems for zebrafish stromal cells derived from organ outgrowths . We prospectively sorted sdf1 DsRed PVC by flow cytometry based on DsRed expression followed by plating onto tissue culture treated plastic dishes.…”

“…hMSCs have the ability to support the expansion of hematopoietic stem and progenitor cells in vitro . We have previously shown that zebrafish stromal cells have a similar ability . We next explored the capacity of the culture‐expanded s df1 DsRed PVC to support hematopoietic cells.…”