Genetic and functional characterization of isolated stromal cell lines from the aorta-gonado-mesonephros region.

University of Tübingen, Department of Hematology, Oncology, and Immunology, Otfried-Müller-Strasse 10, 72076 Tübingen, Germany.
Annals of the New York Academy of Sciences (Impact Factor: 4.31). 07/2005; 1044:51-9. DOI: 10.1196/annals.1349.007
Source: PubMed

ABSTRACT The hematopoietic system interacts with a supportive stromal environment allowing maintenance and differentiation of hematopoietic stem cells (HSCs). The aorta-gonado-mesonephros (AGM) region serves as a unique embryonic microenvironment, generating the first adult repopulating HSCs in the mouse embryo. To eludicate factors involved in hematopoietic support and induction of hematopoietic differentiation, we isolated more than 100 stromal cell clones derived from the AGM region of embryonic day (E) 10.5 mouse embryos for functional and genetic analysis. Selected isolated AGM stromal cell lines are highly efficient in supporting maintenance and expansion of mouse and human hematopoietic stem and progenitor cells. In addition, we can demonstrate for the first time that AGM stromal cell lines are also potent inducers of hematopoietic differentiation of murine embryonic stem cells. Stromal gene array analysis has identified genes that could play a role in hematopoietic support.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: We previously established a mesenchymal stem cell line (FMS/PA6-P) from the bone marrow adherent cells of fetal mice. The cell line expresses a higher level of neural cell adhesion molecule and shows greater hematopoiesis-supporting capacity in mice than other murine stromal cell lines. Since there is 94% homology between human and murine neural cell adhesion molecule, we examined whether FMS/PA6-P cells support human hematopoiesis and whether neural cell adhesion molecules expressed on FMS/PA6-P cells contribute greatly to the human hematopoiesis-supporting ability of the cell line. When lineage-negative cord blood mononuclear cells were co-cultured on the FMS/PA6-P cells, a significantly greater hematopoietic stem cell-enriched population (CD34(+)CD38(-) cells) was obtained than in the culture without the FMS/PA6-P cells. Moreover, when lineage-negative cord blood mononuclear cells were cultured on FMS/PA6-P cells and transplanted into SCID mice, a significantly larger proportion of human CD45(+) cells and CD34(+)CD38(-) cells were detected in the bone marrow of SCID mice than in the bone marrow of SCID mice that had received lineage-negative cord blood mononuclear cells cultured without FMS/PA6-P cells. Furthermore, we found that direct cell-to-cell contact between the lineage-negative cord blood mononuclear cells and the FMS/PA6-P cells was essential for the maximum expansion of the mononuclear cells. The addition of anti-mouse neural cell adhesion molecule antibody to the culture significantly inhibited their contact and the proliferation of lineage-negative cord blood mononuclear cells. These findings suggest that neural cell adhesion molecules expressed on FMS/PA6-P cells play a crucial role in the human hematopoiesis-supporting ability of the cell line.
    Haematologica 12/2009; 95(6):884-91. DOI:10.3324/haematol.2009.013151 · 5.94 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Hematopoietic stem cell (HSC) transplantation is an important treatment modality for hematological malignancies or to correct congenital immunodeficiency disorders. Several stem cell sources are currently applied clinically, with a recent increased application of umbilical cord blood. The low number of HSCs available, particularly in umbilical cord blood, is a limiting factor, and different lines of research are ongoing to circumvent this issue. In this review, we will describe the research strategies developed to expand adult HSCs in vitro and to generate new HSCs from pluripotent stem cell lines. We will also discuss the importance of studying the embryonic microenvironment since it allows both generation and extensive expansion of HSCs. Understanding the mechanisms that underlie HSC production, self-renewal and differentiation is necessary for the establishment of optimal in vitro HSC cultures, where a limitless and manipulatable resource of HSCs would be available for both clinical and fundamental research.
    Regenerative Medicine 05/2012; 7(3):349-68. DOI:10.2217/rme.11.120 · 3.87 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: During mammalian embryonic development the definitive haematopoietic stem cells (HSCs) may arise either in the extra-embryonic mesoderm or in the aorta-gonad-mesonephros (AGM) region that forms in close proximity to the assembling urogenital system, generating the gonad, cortex of the adrenal gland and metanephros. Researchers have been attempting for a long time to define the region of importance for generating the definitive HSCs that colonize the fetal liver and bone marrow, the two major sites where haematopoiesis takes place in the adult. The fetal liver might gain HSCs from both of the primary haematopoietic sources, but the extra-embryonic HSCs seem not to be able to colonize adult bone marrow directly. It is known that the microenvironment around the HSCs is important for directing cell fates, but we do not yet have much idea about the cell-cell interactions, tissue interactions and molecules that regulate cell behaviour in the AGM. We will here discuss the contribution of the AGM to definitive haematopoiesis in mammals and review some of the cell-cell interactions and associated signalling systems involved in the development of AGM stem cells.
    Apmis 11/2005; 113(11-12):804-12. DOI:10.1111/j.1600-0463.2005.apm_368.x · 1.92 Impact Factor


Available from