Erwin W Mark, Ashman Keith, O'Donnel Paul, Inman Robert D
Toronto Western Research Institute, University of Toronto, Toronto, Ontario, Canada.
Arthritis Rheum. 2006 Dec;54(12):3859-67. doi: 10.1002/art.22258.
To identify the components of conditioned medium obtained from intervertebral disc nucleus pulposus-derived canine notochord cells, and to evaluate the capacity of such factors to affect disc-derived chondrocyte gene expression of aggrecan, versican, and hyaluronic acid synthase 2 (HAS-2) as a function of culture conditions.
Canine notochord cells obtained from nonchondrodystrophic dogs were cultured within alginate beads under conditions of serum deficiency (Dulbecco's modified Eagle's medium [DMEM]) to produce notochord cell-conditioned medium (NCCM). NCCM was evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and liquid chromatography-tandem mass spectroscopy. Bovine disc-derived chondrocytes were cultured with serum-deficient medium (DMEM) and NCCM and assayed for the effect of tissue culture conditions on aggrecan, versican, and HAS-2 gene expression. Next, chondrocyte gene expression for aggrecan was evaluated using DMEM containing recombinant connective tissue growth factor (rCTGF), and the results compared with those obtained using NCCM and DMEM.
NCCM contained aggrecan, Cu/Zn superoxide dismutase, fibronectin, and CTGF precursor. Culture with NCCM caused an up-regulation of aggrecan, versican, and HAS-2 gene expression. NCCM induced aggrecan gene expression in chondrocytes at a level similar to that induced by 100-200 ng/ml rCTGF. Nonchondrodystrophic and chondrodystrophic canine notochord cells exhibited similar levels of CTGF gene expression.
Nucleus pulposus-derived notochord cells secrete CTGF (CCN2), a recently discovered multifunctional growth factor. There is no difference between CTGF gene expression in nonchondrodystrophic and chondrodystrophic canine notochord cells, suggesting a possible role of CTGF as an anabolic factor within the disc nucleus that is, to at least some degree, dependent on the population of notochord cells within the disc nucleus.
鉴定从犬椎间盘髓核来源的脊索细胞获得的条件培养基的成分，并评估这些因子在不同培养条件下影响椎间盘来源软骨细胞中聚集蛋白聚糖、多功能蛋白聚糖和透明质酸合酶2（HAS-2）基因表达的能力。
从非软骨发育不良犬获取的犬脊索细胞在血清缺乏条件下（杜氏改良 Eagle 培养基[DMEM]）于藻酸盐珠内培养，以产生脊索细胞条件培养基（NCCM）。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和液相色谱-串联质谱对 NCCM 进行评估。牛椎间盘来源的软骨细胞用血清缺乏培养基（DMEM）和 NCCM 培养，并检测组织培养条件对聚集蛋白聚糖、多功能蛋白聚糖和 HAS-2 基因表达的影响。接下来，使用含重组结缔组织生长因子（rCTGF）的 DMEM 评估聚集蛋白聚糖的软骨细胞基因表达，并将结果与使用 NCCM 和 DMEM 获得的结果进行比较。
NCCM 含有聚集蛋白聚糖、铜/锌超氧化物歧化酶、纤连蛋白和 CTGF 前体。用 NCCM 培养导致聚集蛋白聚糖、多功能蛋白聚糖和 HAS-2 基因表达上调。NCCM 在软骨细胞中诱导的聚集蛋白聚糖基因表达水平与 100 - 200 ng/ml rCTGF 诱导的水平相似。非软骨发育不良和软骨发育不良犬的脊索细胞表现出相似水平的 CTGF 基因表达。
髓核来源的脊索细胞分泌 CTGF（CCN2），一种最近发现的多功能生长因子。非软骨发育不良和软骨发育不良犬的脊索细胞中 CTGF 基因表达无差异，提示 CTGF 作为椎间盘髓核内合成代谢因子可能发挥作用，即至少在一定程度上依赖于椎间盘髓核内脊索细胞群体。
