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      • Record: found
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      Wnt5a Does Not Support Hematopoiesis in Stroma-Free, Serum-Free Cultures

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          Abstract

          Previously we reported that Wnt5a is highly expressed in the murine urogenital ridge-derived UG26-1B6 cells but not embryonic liver-derived EL08-1D2 cells. Mouse long-term repopulating hematopoietic stem cells (LTR-HSC) were maintained in non-contact UG26-1B6 cultures but not EL08-1D2 non-contact cultures, unless Wnt5a was also added to the cultures, suggesting a role for Wnt5a in the in vitro maintenance of LTR-HSC. Here, we investigated if the effect of Wnt5a on adult LTR-HSC activity is HSC-autonomous. To test the effect of Wnt5a on maintenance of LTR-HSC, we performed limiting dilution competitive transplantation assays of murine Lin-Sca1 + c-kit + (LSK) cells cultured for 5 days with TPO and SCF with and without Wnt5a. The effect of Wnt5a on the generation of colony forming units (CFU) and the homing ability of LSK progeny was also tested. No effects were found of Wnt5a on total cell expansion, the number of CFU, or homing ability of day 5 LSK progeny. Furthermore, addition of Wnt5a did not improve, but may have impeded maintenance of LTR-HSC. In conclusion, our data indicate that Wnt5a does not enhance the maintenance and expansion of adult murine LTR-HSCs or committed progenitors cultured in vitro in serum- and stroma-free conditions.

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          Most cited references15

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          A Wnt5a pathway underlies outgrowth of multiple structures in the vertebrate embryo.

          T Yamaguchi, A. Bradley, A McMahon … (1999)
          Morphogenesis depends on the precise control of basic cellular processes such as cell proliferation and differentiation. Wnt5a may regulate these processes since it is expressed in a gradient at the caudal end of the growing embryo during gastrulation, and later in the distal-most aspect of several structures that extend from the body. A loss-of-function mutation of Wnt5a leads to an inability to extend the A-P axis due to a progressive reduction in the size of caudal structures. In the limbs, truncation of the proximal skeleton and absence of distal digits correlates with reduced proliferation of putative progenitor cells within the progress zone. However, expression of progress zone markers, and several genes implicated in distal outgrowth and patterning including Distalless, Hoxd and Fgf family members was not altered. Taken together with the outgrowth defects observed in the developing face, ears and genitals, our data indicates that Wnt5a regulates a pathway common to many structures whose development requires extension from the primary body axis. The reduced number of proliferating cells in both the progress zone and the primitive streak mesoderm suggests that one function of Wnt5a is to regulate the proliferation of progenitor cells.
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            The biology of stem cell factor and its receptor C-kit.

            Leonie Ashman (1999)
            The receptor tyrosine kinase c-Kit and its ligand Stem Cell Factor (SCF) are essential for haemopoiesis, melanogenesis and fertility. SCF acts at multiple levels of the haemopoietic hierarchy to promote cell survival, proliferation, differentiation, adhesion and functional activation. It is of particular importance in the mast cell and erythroid lineages, but also acts on multipotential stem and progenitor cells, megakaryocytes, and a subset of lymphoid progenitors. SCF exists in soluble or transmembrane forms which appear to differ in function. Multiple isoforms of c-Kit also exist as a result of alternate mRNA splicing, proteolytic cleavage and the use of cryptic internal promoters in certain cell types. This review focuses on what is known about the regulation of c-Kit expression, the functions of SCF and c-Kit isoforms, and the nature of the biological responses elicited by this receptor-ligand pair with emphasis on the haemopoietic system.
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              Wnt signaling regulates B lymphocyte proliferation through a LEF-1 dependent mechanism.

              R Nusse, Stephen O’Riordan, E Devaney … (2000)
              Lymphocyte enhancer factor-1 (LEF-1) is a member of the LEF-1/TCF family of transcription factors, which have been implicated in Wnt signaling and tumorigenesis. LEF-1 was originally identified in pre-B and T cells, but its function in B lymphocyte development remains unknown. Here we report that LEF-1-deficient mice exhibit defects in pro-B cell proliferation and survival in vitro and in vivo. We further show that Lef1-/- pro-B cells display elevated levels of fas and c-myc transcription, providing a potential mechanism for their increased sensitivity to apoptosis. Finally, we establish a link between Wnt signaling and normal B cell development by demonstrating that Wnt proteins are mitogenic for pro-B cells and that this effect is mediated by LEF-1.
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                Author and article information

                Contributors
                Zoran Ivanovic: Role: Editor
                Journal
                Journal ID (nlm-ta): PLoS One
                Journal ID (iso-abbrev): PLoS ONE
                Journal ID (publisher-id): plos
                Journal ID (pmc): plosone
                Title: PLoS ONE
                Publisher: Public Library of Science (San Francisco, USA )
                ISSN (Electronic): 1932-6203
                Publication date Collection: 2013
                Publication date (Electronic): 14 January 2013
                Volume: 8
                Issue: 1
                Electronic Location Identifier: e53669
                Affiliations
                [1]Interdepartementeel Stamcelinstituut, Katholieke Universiteit Leuven, Leuven, Belgium
                French Blood Institute, France
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: AMSO SS CMV. Performed the experiments: AMSO SS SK. Analyzed the data: AMSO SS CMV. Contributed reagents/materials/analysis tools: AMSO SS SK CMV. Wrote the paper: AMSO CMV.

                Article
                Publisher ID: PONE-D-12-18327
                DOI: 10.1371/journal.pone.0053669
                PMC ID: 3544900
                PubMed ID: 23341970
                SO-VID: b90acec2-111e-439c-8e6b-0e7bdb0f0980
                Copyright statement: Copyright @ 2013
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                Date received : 24 June 2012
                Date accepted : 3 December 2012
                Page count
                Pages: 6
                Funding
                This study was supported by FWO funding (G085111N), NIH-PO1-CA-65493-06, Odysseus funding, CoE and GOA/11/012 funding from K.U.Leuven, and the Vanwayenberghe fund to CM Verfaillie. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Subject: Research Article
                Subject: Biology
                Subject: Developmental Biology
                Subject: Stem Cells
                Subject: Adult Stem Cells
                Subject: Hematopoietic Stem Cells
                Subject: Model Organisms
                Subject: Animal Models
                Subject: Mouse
                Subject: Molecular Cell Biology
                Subject: Signal Transduction
                Subject: Signaling Cascades
                Subject: WNT Signaling Cascade

                ScienceOpen disciplines: Uncategorized
                Data availability:
                ScienceOpen disciplines: Uncategorized

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