Biochemical characterization of heparan sulfate derived from murine hemopoietic stromal cell lines: A bone marrow‐derived cell line S17 and a fetal liver‐derived cell line AFT024

Abstract

Heparan sulfate (HS) present on the surface of hemopoietic stromal cells has important roles in the control of adhesion and growth of hemopoietic stem and progenitor cells. Recent studies have characterized several different heparan sulfate proteoglycans (HSPGs) from both human and murine bone marrow stromal cells. In the present study, we have compared the molecular structure of HS, metabolically labeled with [(35)S]-sulfate produced by two distinct preparations of murine hemopoietic stromal cell lines. These comprised a bone marrow-derived cell line S17 and a fetal liver-derived cell line AFT024. [(35)S]-HS was examined in the cell layers and in the culture medium. We identified and measured the relative proportions of the various glycosaminoglycans (GAGs) in the two stromal cell lines. Chondroitin sulfate (CS) was preponderantly secreted by the stromal cell lines, while HS was relatively more abundant in the cell-associated fractions. The two types of stromal cells differ in their HS composition, mainly due to different patterns of N- and O-sulfation. The two stromal cell lines expressed mRNA for different HSPGs. Data from reverse transcription PCR revealed that the two stromal cell lines expressed mRNA for glypican and syndecan4. Only AFT024 cell line expressed mRNA for betaglycan. There was no evidence for expression of mRNA for both syndecan1 and syndecan2. [(35)S]-sulfated macromolecules could be released from the cell surface of both stromal cell lines by phosphatidylinositol phospholipase C (PI-PLC), which is consistent with the expression of glypican detected by PCR experiments.

Gangliosides of myelosupportive stroma cells are transferred to myeloid progenitors and are required for their survival and proliferation

“…FDC-P1 cells proliferated both in co-culture with AFT-024 stroma ( Figures 1A and 2) and in the presence of the AFT-024 cellculture supernatant ( Figure 1B). This is in accordance with our previous study [29] showing that AFT-024 cells, although lacking IL-3, produced and secreted high amounts GM-CSF, as well as heparan-sulphate-bearing proteoglycans that interacted with GM-CSF. These stroma cells could thus be used for studies on the GM-CSF-dependent myeloid cell proliferation.…”

Stroma-mediated granulocyte-macrophage colony-stimulating factor (GM-CSF) control of myelopoiesis: spatial organisation of intercellular interactions

“…Proteoglycans can be transmembrane molecules or anchored in the external membrane leaflet, and their mobility in the membrane can be selectively controlled. We have already shown that this property can be determinant for the proteoglycan participation in controls of the stroma-mediated myelopoiesis (Arcanjo et al 2002). Taken together, our results indicate that the specificity of stroma-dependent controls of a given growth factor's activity depends upon a combinatorial interplay among the cytokine, the quality of membrane-associated molecules and the spatial organisation of the intercellular environment.…”

Efficient Generation of Plasmacytoid Dendritic Cell from Common Lymphoid Progenitors by Flt3 Ligand

“…It is known that a stromal cell line can constitutively produce hematopoietic cytokines such as IL-7, SCF, M-CSF and FL to support hematopoiesis in vitro [ 35 ]. Moreover, heparan sulfate present on the surface of hematopoietic stromal cells has been shown to control adhesion and growth of hematopoietic stem and progenitor cells [ 36 ]. Therefore, differences in quantity or quality of these factors in FF, BF and AC-6 system may account for the distinct phenotypes.…”