Drzeniek Z, Stöcker G, Siebertz B, Just U, Schroeder T, Ostertag W, Haubeck H D
Institute for Clinical Chemistry and Pathobiochemistry, Medical Faculty, University of Technology, Aachen, Germany.
Blood. 1999 May 1;93(9):2884-97.
Heparan sulfate (HS) proteoglycans of bone marrow (BM) stromal cells and their extracellular matrix are important components of the microenvironment of hematopoietic tissues and are involved in the interaction of hematopoietic stem and stromal cells. Although previous studies have emphasized the role of HS proteoglycan synthesis by BM stromal cells, we have recently shown that the human hematopoietic progenitor cell line TF-1 also expressed an HS proteoglycan. Immunochemical, reverse transcriptase-polymerase chain reaction (RT-PCR), and Northern blot analysis of this HS proteoglycan showed that it was not related to the syndecan family of HS proteoglycans or to glypican. To answer the question of whether the expression of HS proteoglycans is associated with the differentiation state of hematopoietic progenitor cells, we have analyzed the proteoglycan synthesis of several murine and human hematopoietic progenitor cell lines. Proteoglycans were isolated from metabolically labeled cells and purified by several chromatographic steps. Isolation and characterization of proteoglycans from the cell lines HEL and ELM-D, which like TF-1 cells have an immature erythroid phenotype, showed that these cells synthesize the same HS proteoglycan, previously detected in TF-1 cells, as a major proteoglycan. In contrast, cell lines of the myeloid lineage, like the myeloblastic/promyelocytic cell lines B1 and B2, do not express HS proteoglycans. Taken together, our data strongly suggest that expression of this HS proteoglycan in hematopoietic progenitor cell lines is associated with the erythroid lineage. To prove this association we have analyzed the proteoglycan expression in the nonleukemic multipotent stem cell line FDCP-Mix-A4 after induction of erythroid or granulocytic differentiation. Our data show that HS proteoglycan expression is induced during early erythroid differentiation of multipotent hematopoietic stem cells. In contrast, during granulocytic differentiation, no expression of HS proteoglycans was observed.
骨髓（BM）基质细胞及其细胞外基质中的硫酸乙酰肝素（HS）蛋白聚糖是造血组织微环境的重要组成部分，参与造血干细胞与基质细胞的相互作用。尽管先前的研究强调了BM基质细胞合成HS蛋白聚糖的作用，但我们最近发现人类造血祖细胞系TF-1也表达一种HS蛋白聚糖。对这种HS蛋白聚糖进行免疫化学、逆转录聚合酶链反应（RT-PCR）和Northern印迹分析表明，它与HS蛋白聚糖的syndecan家族或glypican无关。为了回答HS蛋白聚糖的表达是否与造血祖细胞的分化状态相关这一问题，我们分析了几种小鼠和人类造血祖细胞系的蛋白聚糖合成情况。从代谢标记的细胞中分离蛋白聚糖，并通过几个色谱步骤进行纯化。从与TF-1细胞一样具有未成熟红系表型的细胞系HEL和ELM-D中分离和鉴定蛋白聚糖，结果表明这些细胞合成的主要蛋白聚糖与先前在TF-1细胞中检测到的HS蛋白聚糖相同。相反髓系细胞系，如髓母细胞/早幼粒细胞系B1和B2，不表达HS蛋白聚糖。综上所述，我们的数据强烈表明造血祖细胞系中这种HS蛋白聚糖的表达与红系谱系相关。为了证明这种关联，我们分析了非白血病多能干细胞系FDCP-Mix-A4在诱导红系或粒系分化后的蛋白聚糖表达情况。我们的数据表明，多能造血干细胞在早期红系分化过程中诱导HS蛋白聚糖表达。相反，在粒系分化过程中，未观察到HS蛋白聚糖的表达。
