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.
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.
Arcanjo
Katia
K
Departamento de Histologia e Embriologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Belo
Gisele
G
Folco
Cristiane
C
Werneck
Claudio C
CC
Borojevic
Radovan
R
Silva
Luiz-Claudio F
LC
D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism*; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000378:metabolism; D002808:Chondroitin Lyases / Q000737:chemistry / Q000378:metabolism; D002852:Chromatography, Ion Exchange / Q000379:methods; D016207:Cytokines / Q000096:biosynthesis; D017931:DNA Primers; D004187:Disaccharides / Q000096:biosynthesis / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006493:Heparin / Q000737:chemistry / Q000378:metabolism; D006497:Heparan Sulfate / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D009608:Nitrous Acid / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000378:metabolism; D013462:Sulfur Radioisotopes; D013997:Time Factors
D002808:Chondroitin Lyases; D016207:Cytokines; D017931:DNA Primers; D004187:Disaccharides; D006493:Heparin; D006497:Heparan Sulfate; D009608:Nitrous Acid; D013462:Sulfur Radioisotopes