Proteoglycan synthesis in haematopoietic cells: isolation and characterization of heparan sulphate proteoglycans expressed by the bone-marrow stromal cell line MS-5

Drzeniek, Zofia; Siebertz, Barbara; Stöcker, Georg; Just, Ursula; Ostertag, Wolfram; Greiling, H.; Haubeck, H.-D.

doi:10.1042/bj3270473

Cited by 33 publications

(28 citation statements)

References 62 publications

(68 reference statements)

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“…Our results suggest that syndecan-4, syndecan-3 and glypican-1, present on the surface of marrow stromal cells, together with perlecan in the ECM, may be responsible for creating the correct stromal niche for the maintenance and development of haemopoietic stem and progenitor cells. These findings differ from those of a recent study of a murine stromal cell line in which syndecan-1 and syndecan-2 were the most abundant HSPGs, followed by syndecan-3 and syndecan-4 [15]. It is possible that the differences in human and murine expression of proteoglycans may relate to the ability of human stromas to more efficiently maintain haemopoietic stem cells.…”

Section: Discussioncontrasting

confidence: 95%

“…The marrow environment in i o is mimicked by the adherent stromal layer that forms in long-term culture which is able to support the growth of haemopoietic progenitor cells [1]. A recent study has characterized several different HSPGs from the murine bone marrow stromal cell line MS-5 that can support the growth of human progenitor cells [15], but although the molecular structure of the glycosaminoglycan chains of heparan sulphate has been analysed in human long-term bone marrow cultures [16], the associated proteoglycan core proteins have hitherto been uncharacterized.…”

Section: Introductionmentioning

confidence: 99%

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Expression of proteoglycan core proteins in human bone marrow stroma

Schofield

Gallagher

Gourichon

1999

Biochemical Journal

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Heparan sulphate proteoglycans (HSPGs) present on the surface of bone marrow stromal cells and in the extracellular matrix (ECM) have important roles in the control of adhesion and growth of haemopoietic stem and progenitor cells. The two main groups of proteoglycans which contain heparan sulphate chains are members of the syndecan and glypican families. In this study we have identified the main surface membrane and matrixassociated HSPGs present in normal human bone marrow stroma formed in long-term culture. Proteoglycans were extracted from the adherent stromal layers and treated with heparitinase and chondroitinase ABC. The core proteins were detected by Western blotting using antibodies directed against syndecans-1-4, glypican-1 and the ECM HSPG, perlecan. Stromal cell expression at the RNA level was detected by Northern blotting and by reverse transcription PCR. Glypican-1, syndecan-3 and syn-

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Section: Discussioncontrasting

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Expression of proteoglycan core proteins in human bone marrow stroma

Schofield

Gallagher

Gourichon

1999

Biochemical Journal

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“…However, no GAGs were detected in HUVEC monoculture (Fig. 4) Interestingly, it is thought that heparan sulfate proteoglycans assist in mediating the interactions between marrow stromal cells and hematopoietic cells, 42,43 and thus, the upregulation of GAG production observed in the present study potentially could have further enhanced the effect of the cocultures.…”

Section: Discussionmentioning

confidence: 69%

Enhanced Osteogenesis in Cocultures with Human Mesenchymal Stem Cells and Endothelial Cells on Polymeric Microfiber Scaffolds

Gershovich

Dahlin

Kasper

et al. 2013

Tissue Engineering Part A

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In this work, human mesenchymal stem cells (hMSCs) and their osteogenically precultured derivatives were directly cocultured with human umbilical vein endothelial cells (HUVECs) on electrospun three-dimensional poly(e-caprolactone) microfiber scaffolds to evaluate the coculture's effect on the generation of osteogenic constructs. Specifically, cells were cultured on scaffolds for up to 3 weeks, and the cellularity, alkaline phosphatase (ALP) activity, and bone-like matrix formation were assessed. Constructs with cocultures and monocultures had almost identical cellularity after the first week, however, lower cellularity was observed in cocultures compared to monocultures during the subsequent 2 weeks of culture. Scaffolds with cocultures showed a significantly higher ALP activity, glycosaminoglycan and collagen production, as well as greater calcium deposition over the course of study compared to monocultures of hMSCs. Furthermore, the osteogenic outcome was equally robust in cocultures containing osteogenically precultured and non-precultured hMSCs. The results demonstrate that the combination of MSC and HUVEC populations within a porous scaffold material under osteogenic culture conditions is an effective strategy to promote osteogenesis.

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“…However, our knowledge on the heparan sulphate proteoglycans, involved in the interaction of haematopoietic-progenitor and stromal cell lines, is still limited. In fact, there are only few reports on proteoglycans from haematopoietic-bone-marrow cells [24,[37][38][39][40][41][42][43][44] pointing to stromal cells, rather than haematopoietic-stem cells or progenitor cells, as the source of heparan sulphate proteoglycans [24,[37][38][39][40][42][43][44]. Recently, we have analysed the expression of heparan sulphate proteoglycans in several murine and human haematopoietic-progenitor and stromal cell lines [43].…”

Section: Discussionmentioning

confidence: 99%

“…In fact, there are only few reports on proteoglycans from haematopoietic-bone-marrow cells [24,[37][38][39][40][41][42][43][44] pointing to stromal cells, rather than haematopoietic-stem cells or progenitor cells, as the source of heparan sulphate proteoglycans [24,[37][38][39][40][42][43][44]. Recently, we have analysed the expression of heparan sulphate proteoglycans in several murine and human haematopoietic-progenitor and stromal cell lines [43]. In contrast with the bone-marrow-stromal cell line MS-5, which expressed at least seven different heparan sulphate proteoglycans, the expression of heparan sulphate proteoglycans in haematopoieticprogenitor cell lines is limited [35,43].…”

Section: Discussionmentioning

confidence: 99%

Expression of glypican-4 in haematopoietic-progenitor and bone-marrow-stromal cells

Siebertz

Stöcker

Drzeniek

et al. 1999

Biochemical Journal

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Heparan sulphate proteoglycans and the extracellular matrix of bone-marrow-stromal cells are important components of the microenvironment of haematopoietic tissues and are involved in the interaction of haematopoietic stem and stromal cells. Previous studies have emphasized the role of heparan sulphate proteoglycan synthesis by bone-marrow-stromal cells. In the present study we describe the expression of glypican-4 (GPC-4), belonging to the glypican family, in bone-marrow-stromal cells and haematopoietic-progenitor cells of human and murine origin. Expression of GPC-4 was shown on the mRNA-level by reverse transcription-PCR and Northern blot analysis. Amplification products were cloned and sequenced, to confirm these results. To analyze the expression of GPC-4 on the protein level, polyclonal antibodies against selected peptides were raised in rabbits. Western blot analysis showed expression of GPC-4 as a heparan sulphate proteoglycan in the human haematopoietic-progenitor cell line TF-1 and normal human bone marrow. These results were confirmed by FACS analysis of TF-1 cells. Furthermore, GPC-4-positive progenitor cells and stromal cells were enriched from normal human bone marrow by magnetic-cell sorting and analysed by confocal laser-scanning microscopy.

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Proteoglycan synthesis in haematopoietic cells: isolation and characterization of heparan sulphate proteoglycans expressed by the bone-marrow stromal cell line MS-5

Cited by 33 publications

References 62 publications

Expression of proteoglycan core proteins in human bone marrow stroma

Expression of proteoglycan core proteins in human bone marrow stroma

Enhanced Osteogenesis in Cocultures with Human Mesenchymal Stem Cells and Endothelial Cells on Polymeric Microfiber Scaffolds

Expression of glypican-4 in haematopoietic-progenitor and bone-marrow-stromal cells

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