Notochordal cell conditioned medium stimulates mesenchymal stem cell differentiation toward a young nucleus pulposus phenotype.
Notochordal cell conditioned medium stimulates mesenchymal stem cell differentiation toward a young nucleus pulposus phenotype.
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DOI：
10.1186/scrt18
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发表时间：
2010-06-16
影响因子：
7.5
通讯作者：
Iatridis JC
中科院分区：
文献类型：
--
作者：
Korecki CL;Taboas JM;Tuan RS;Iatridis JC
间充质干细胞（MSC）为椎间盘（IVD）修复和再生提供了希望，因为它们易于分离和扩增，并且可以分化成多种间充质组织。脊索（NC）细胞有助于IVD的发展，纳入髓核（NP），并刺激成熟的椎间盘细胞。然而，还没有研究调查的影响，NC细胞对成体干细胞分化。本研究的前提是，IVD再生更类似于IVD的发展比IVD的维护，我们假设，可溶性因子从NC细胞分化间充质干细胞的髓核（NP）细胞的表型特征在发展过程中。最终的临床目标将是分离或化学/重组产生活性剂以诱导治疗效果，并将其用作用于椎间盘疾病早期干预的可注射疗法，或用于在组织工程化NP构建体中开发适当预分化的MSC细胞。将来自骨髓的人MSC扩增并沉淀以形成高密度培养物。将MSC团块暴露于对照培养基（CM）、软骨形成培养基（含有地塞米松和转化生长因子（TGF）-β3的CM）或脊索细胞条件培养基（NCCM）。从在无血清培养基中维持4天的NC细胞制备NCCM。培养7天后，分析MSC团块的外观、生化组成（糖胺聚糖和DNA）和基因表达谱（sox-9、II型和III型胶原、层粘连蛋白-β1和TIMP 1（金属蛋白酶-1的组织抑制剂））。在NCCM处理的团块中观察到比CM或TGFβ组显著更高的糖胺聚糖积累。与TGFβ组相比，NCCM组的III型胶原基因表达增加，层粘连蛋白β1表达增加，sox-9和II型胶原表达减少。总之，结果表明NCCM刺激间充质干细胞分化为潜在的NP样表型，具有发展中IVD的一些特征。
Mesenchymal stem cells (MSCs) offer promise for intervertebral disc (IVD) repair and regeneration because they are easily isolated and expanded, and can differentiate into several mesenchymal tissues. Notochordal (NC) cells contribute to IVD development, incorporate into the nucleus pulposus (NP), and stimulate mature disc cells. However, there have been no studies investigating the effects of NC cells on adult stem cell differentiation. The premise of this study is that IVD regeneration is more similar to IVD development than to IVD maintenance, and we hypothesize that soluble factors from NC cells differentiate MSCs to a phenotype characteristic of nucleus pulposus (NP) cells during development. The eventual clinical goal would be to isolate or chemically/recombinantly produce the active agent to induce the therapeutic effects, and to use it as either an injectable therapy for early intervention on disc disease, or in developing appropriately pre-differentiated MSC cells in a tissue engineered NP construct. Human MSCs from bone marrow were expanded and pelleted to form high-density cultures. MSC pellets were exposed to either control medium (CM), chondrogenic medium (CM with dexamethasone and transforming growth factor, (TGF)-β3) or notochordal cell conditioned medium (NCCM). NCCM was prepared from NC cells maintained in serum free medium for four days. After seven days culture, MSC pellets were analyzed for appearance, biochemical composition (glycosaminoglycans and DNA), and gene expression profile (sox-9, collagen types-II and III, laminin-β1 and TIMP1(tissue inhibitor of metalloproteinases-1)). Significantly higher glycosaminoglycan accumulation was seen in NCCM treated pellets than in CM or TGFβ groups. With NCCM treatment, increased gene expression of collagen III, and a trend of increasing expression of laminin-β1 and decreased expression of sox-9 and collagen II relative to the TGFβ group was observed. Together, results suggest NCCM stimulates mesenchymal stem cell differentiation toward a potentially NP-like phenotype with some characteristics of the developing IVD.
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影响因子：
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作者：
Erwin, W. Mark;Ashman, Keith;Inman, Robert D.
通讯作者：
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影响因子：
17.3
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