title: Proteoglycan synthesis in haematopoietic cells: isolation and characterization of heparan sulphate proteoglycans expressed by the bone-marrow stromal cell line MS-5.
abstract: Proteoglycans of bone-marrow stromal cells and their extracellular matrix are important components of the haematopoietic microenvironment. Recently, several studies have indicated that they are involved in the interaction of haematopoietic stem and stromal cells. However, a detailed characterization of the heparan sulphate proteoglycans synthesized by bone-marrow stromal cells is still lacking. Here we report on the isolation and characterization of proteoglycans from the haematopoietic stromal cell line MS-5, that efficiently supports the growth and differentiation of human and murine haematopoietic progenitor cells. Biochemical characterization of purified proteoglycans revealed that the haematopoietic stromal cell line MS-5 synthesizes, in addition to chondroitin sulphate proteoglycans, several different heparan sulphate proteoglycans. Immunochemical analysis, using specific antibodies against the different members of the syndecan family, glypican, betaglycan and perlecan, showed that MS-5 cells synthesize all these different heparan sulphate proteoglycans. These data were further supported by reverse-transcriptase PCR and confirmed by sequence and Northern blot analysis. The relative abundance of the different heparan sulphate proteoglycans was estimated on the protein and mRNA levels.
authors.affiliation: Institute for Clinical Chemistry and Pathobiochemistry, Medical Faculty, University of Technology, Pauwelsstr. 30, D-52057 Aachen, Germany.
authors.forename: Z
authors.initials: Z
authors.lastname: Drzeniek
authors.forename: B
authors.initials: B
authors.lastname: Siebertz
authors.forename: G
authors.initials: G
authors.lastname: Stöcker
authors.forename: U
authors.initials: U
authors.lastname: Just
authors.forename: W
authors.initials: W
authors.lastname: Ostertag
authors.forename: H
authors.initials: H
authors.lastname: Greiling
authors.forename: H D
authors.initials: HD
authors.lastname: Haubeck
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation; D002455:Cell Division; D002460:Cell Line; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D019812:Heparan Sulfate Proteoglycans / Q000096:biosynthesis*; D006801:Humans; D051379:Mice; D016133:Polymerase Chain Reaction; D011509:Proteoglycans / Q000096:biosynthesis; D018125:Receptors, Transforming Growth Factor beta / Q000096:biosynthesis; D017154:Stromal Cells / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism
chemical_list: D019812:Heparan Sulfate Proteoglycans; D011509:Proteoglycans; D018125:Receptors, Transforming Growth Factor beta; D013431:Sulfates; C071736:betaglycan