Rejuvenation of Nucleus Pulposus Cells Using Extracellular Matrix Deposited by Synovium-Derived Stem Cells
Rejuvenation of Nucleus Pulposus Cells Using Extracellular Matrix Deposited by Synovium-Derived Stem Cells
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DOI：
10.1097/brs.0b013e31821fcc64
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发表时间：
2012-03-15
期刊：
影响因子：
3
通讯作者：
Pei, Ming
中科院分区：
文献类型：
--
作者：
He, Fan;Pei, Ming
研究设计。在塑料瓶或滑膜源性干细胞 (SDSC) 沉积的细胞外基质 (ECM) 上铺板 6 代后，评估扩增的髓核 (NP) 细胞的再分化能力。 目的。目的是评估使用组织特异性干细胞沉积的 ECM 为 NP 细胞再生提供 3 维微环境的可行性。背景数据摘要。基于自体椎间盘细胞的疗法是椎间盘再生的一种有前途的方法。不幸的是，目前单层 NP 细胞的体外扩增导致这些细胞去分化。方法。将原代 NP 细胞铺板在塑料瓶或 ECM 上连续 6 代。在每次传代时，对细胞数量进行增殖率计数，使用流式细胞术评估细胞表型，并使用实时聚合酶链反应（PCR）评估细胞分化状态。将第 1、4 和 6 代扩增的 NP 细胞沉淀在无血清成分确定的培养基中孵育 14 天。使用组织学、生物化学和实时 PCR 评估扩增的 NP 细胞的再分化能力。结果。与塑料瓶上的相比，在 ECM 上扩增的 NP 细胞生长速度更快，尺寸更小，形状类似成纤维细胞。 ECM 处理的 NP 细胞获得了增强的 CD90 表达和更高的 I、II 和 X 型胶原蛋白和聚集蛋白聚糖的 mRNA 水平，以及强大的再分化能力，通过最多 6 代的 II 型胶原蛋白、聚集蛋白聚糖和 Sox9 显着增加以及 I 型胶原蛋白减少来证明。结论。 SDSC衍生的ECM可以为具有更高增殖率和再分化能力的NP细胞的再生提供组织特异性微环境。这些特征可能在改善临床环境中基于自体椎间盘细胞的微创治疗方法以实现生物功能椎间盘的生理重建方面发挥作用。
Study Design. After plating for 6 passages on either plastic flasks or extracellular matrix (ECM) deposited by synovium-derived stem cells (SDSCs), expanded nucleus pulposus (NP) cells were evaluated for redifferentiation capacity.Objective. The aim was to assess the feasibility of using ECM deposited by a tissue-specific stem cell to provide a 3-dimensional microenvironment for NP cell rejuvenation.Summary of Background Data. Autologous disc cell-based therapy is a promising approach for intervertebral disc regeneration. Unfortunately, the current in vitro expansion of NP cells in monolayer results in dedifferentiation of these cells.Methods. Primary NP cells were plated on either plastic flasks or ECM for 6 consecutive passages. At each passage, cell numbers were counted for proliferation rate, cell phenotype was evaluated using flow cytometry, and cell differentiation status was assessed using real-time polymerase chain reaction (PCR). The pellets from expanded NP cells at passages 1, 4, and 6 were incubated in a serum-free defined medium for 14 days. Redifferentiation capacity of the expanded NP cells was evaluated using histology, biochemistry, and real-time PCR.Results. NP cells expanded on ECM grew much faster with a smaller size and fibroblast-like shape compared with those on plastic flasks. ECM-treated NP cells acquired an enhanced CD90 expression and higher mRNA levels of types I, II, and X collagen and aggrecan, as well as a robust redifferentiation capacity, evidenced by dramatically increased type II collagen, aggrecan, and Sox9 and decreased type I collagen for up to 6 passages.Conclusion. SDSC-derived ECM can provide a tissue-specific microenvironment for the rejuvenation of NP cells with a higher proliferation rate and redifferentiation capacity. These characteristics may play a role in improving an autologous disc cell-based minimally invasive therapeutic approach toward physiological reconstruction of a biologically functional disc in the clinical setting.