Influence of Hypoxia in the Intervertebral Disc on the Biological Behaviors of Rat Adipose- and Nucleus Pulposus-Derived Mesenchymal Stem Cells
Influence of Hypoxia in the Intervertebral Disc on the Biological Behaviors of Rat Adipose- and Nucleus Pulposus-Derived Mesenchymal Stem Cells
复制标题
椎间盘缺氧对大鼠脂肪和髓核来源间充质干细胞生物学行为的影响
DOI：
10.1159/000356505
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发表时间：
2013-01-01
影响因子：
2.7
通讯作者：
Chen, Qixin
中科院分区：
文献类型：
--
作者：
Li, Hao;Tao, Yiqing;Chen, Qixin
脂肪来源的间充质干细胞（ADMSC）和髓核来源的间充质干细胞（NPMSC）是用于椎间盘（IVD）再生的基于细胞的疗法的两种细胞候选物。然而，很少有工作已经做了，以确定缺氧的影响，在IVD的ADMSCs和NPMSCs的生物学行为。本研究旨在探讨体外培养的大鼠ADMSCs和NPMSCs在IVD低氧环境中的存活、增殖和分化情况。将分离自6只SD大鼠的ADMSCs和NPMSCs在模拟IVD标准条件和低氧环境的常氧（20%O2）和低氧（2%O2）下培养14天。Annexin-V-FITC/propidium iodide双染法检测细胞活力，MTT法检测细胞增殖。RT-PCR检测缺氧诱导因子-1 α、葡萄糖转运蛋白（GLUT）-1、GLUT-3和血管内皮生长因子-A mRNA水平的表达。在三维微团和分化培养基中培养的细胞中，通过RT-PCR和Western blot检测聚集蛋白聚糖、胶原-II和Sox-9在mRNA和蛋白水平上的表达。缺氧抑制ADMSCs和NPMSCs的活力和增殖，但促进ADMSCs和NPMSCs向软骨细胞分化。与ADMSCs相比，NPMSCs在缺氧条件下表现出更高的活力、增殖和软骨细胞分化。总之，IVD中的缺氧对ADMSC和NPMSC的活力、增殖和软骨细胞分化具有显著影响。在IVD的缺氧环境中，NPMSC表现出比ADMSC更有效的生物活性，并且可能代表用于IVD再生的基于细胞的疗法的另一候选者。
Adipose-derived mesenchymal stem cells (ADMSCs) and nucleus pulposus-derived mesenchymal stem cells (NPMSCs) are two cell candidates for cell-based therapies for intervertebral disc (IVD) regeneration. However, little work has been done to determine the influence of hypoxia in the IVD on the biological behaviors of ADMSCs and NPMSCs. This study aimed to investigate the viability, proliferation and differentiation of rat ADMSCs and NPMSCs in the hypoxic environment of IVD in vitro. ADMSCs and NPMSCs isolated from 6 SD rats were cultured under normoxia (20% O2) and hypoxia (2% O2) mimicking the standard condition and hypoxic environment of the IVD for 14 days. Cell viability was determined by the annexin-V-FITC/propidium iodide double-staining assay and cell proliferation was measured by MTT assay. The expression of hypoxia-inducible factor-1α, glucose transporter (GLUT)-1, GLUT-3 and vascular endothelial growth factor-A at the mRNA level was examined by RT-PCR. In cells cultured in three-dimensional micromass and differentiation medium, aggrecan, collagen-II and Sox-9 expression at mRNA and protein levels were examined by RT-PCR and Western blot. Hypoxia inhibited the viability and proliferation of both ADMSCs and NPMSCs, but promoted the chondrocytic differentiation of ADMSCs and NPMSCs. Compared to ADMSCs, NPMSCs showed greater viability, proliferation and chondrocytic differentiation under hypoxia. In conclusion, hypoxia in the IVD had a significant impact on the viability, proliferation and chondrocytic differentiation of ADMSCs and NPMSCs. NPMSCs exhibited more potent biological activity than ADMSCs in the hypoxic environment of the IVD and may represent another candidate for cell-based therapy for IVD regeneration.