Proteoglycan synthesis in two murine bone marrow stromal cell lines.
Proteoglycan synthesis in two murine bone marrow stromal cell lines.
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DOI：
10.1182/blood.v70.6.1777.1777
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发表时间：
1987-12
期刊：
影响因子：
20.3
通讯作者：
S. Kirby;S. Bentley
中科院分区：
文献类型：
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作者：
S. Kirby;S. Bentley
有证据表明基质蛋白多糖是造血微环境的重要功能成分。因此，在 MS3-2A 和 D2XRII 造血基质细胞系中研究了蛋白多糖的合成。这些细胞系支持体外造血的能力不同，D2XRII 支持体外造血，而 MS3-2A 则不然。用35S-硫酸盐作为前体标记细胞，并通过离子交换色谱、氯化铯密度梯度离心和分子筛色谱对细胞和培养基的4mol/L盐酸胍提取物进行分析。通过酶消化和化学消化进一步检查蛋白多糖。 MS3-2A 细胞产生至少三种蛋白聚糖。两种软骨素/硫酸皮肤素 (CS/DS) 蛋白聚糖，Sepharose CL-2B 上的 Kav = 0.40 和 Kav = 0.68，主要存在于培养基中。各自的糖胺聚糖分子量(mol wt)值为38kd和40kd。 Kav = 0.58 的硫酸乙酰肝素 (HS) 蛋白聚糖和摩尔重量 36 kd 的糖胺聚糖主要存在于细胞层提取物中。 D2XRII 细胞合成两种 HS 蛋白聚糖。较大的（Kav = 0.45；糖胺聚糖摩尔重量，30 kd）在梯度离心时密度较低，在细胞层提取物中更为突出，而较小的（Kav = 0.68；糖胺聚糖摩尔重量，38 kd）密度较高，主要存在于培养基中。 Kav 0.78 的单一 CS/DS 蛋白聚糖种类和摩尔重量 18 kd 的平均糖胺聚糖以大致等量存在于培养基和细胞层中。因此，MS3-2A 和 D2XRII 在蛋白聚糖合成方面表现出不同的表型。这些差异与骨髓基质成分的微环境功能相关。
There is evidence indicating that stromal proteoglycans are an important functional component of the hematopoietic microenvironment. Proteoglycan synthesis was therefore investigated in the MS3-2A and D2XRII hematopoietic stromal cell lines. These lines differ in their capacity to support hematopoiesis in vitro, D2XRII supporting in vitro hematopoiesis, whereas MS3-2A does not. Cells were labeled with 35S-sulfate as precursor, and 4 mol/L guanidine HCl extracts of cells and media were analyzed by ion-exchange chromatography, cesium chloride density gradient centrifugation, and molecular sieve chromatography. Proteoglycans were further examined by enzymatic and chemical digestions. MS3-2A cells produced at least three proteoglycan species. Two chondroitin/dermatan sulfate (CS/DS) proteoglycans, Kav = 0.40 and Kav = 0.68 on Sepharose CL-2B, were present primarily in the medium. The respective glycosaminoglycan molecular weight (mol wt) values were 38 kd and 40 kd. A heparan sulfate (HS) proteoglycan of Kav = 0.58 and glycosaminoglycan mol wt 36 kd was present primarily in the cell layer extract. D2XRII cells synthesized two HS proteoglycans. The larger (Kav = 0.45; glycosaminoglycan mol wt, 30 kd) was of low density on gradient centrifugation and more prominent in the cell layer extracts, whereas the smaller (Kav = 0.68; glycosaminoglycan mol wt, 38 kd) was dense and present mainly in the culture medium. A single CS/DS proteoglycan species of Kav 0.78 and average glycosaminoglycan of mol wt 18 kd was present in roughly equal amounts in the medium and in the cell layer. MS3-2A and D2XRII thus appear phenotypically distinct with respect to proteoglycan synthesis. These differences are discussed in relation to the microenvironmental function of bone marrow stromal elements.