Nucleus Pulposus Mesenchymal Stem Cells in Acidic Conditions Mimicking Degenerative Intervertebral Discs Give Better Performance than Adipose Tissue-Derived Mesenchymal Stem Cells
Nucleus Pulposus Mesenchymal Stem Cells in Acidic Conditions Mimicking Degenerative Intervertebral Discs Give Better Performance than Adipose Tissue-Derived Mesenchymal Stem Cells
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DOI：
10.1159/000369452
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发表时间：
2014-01-01
影响因子：
2.7
通讯作者：
Chen, Qi-xin
中科院分区：
文献类型：
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作者：
Han, Bin;Wang, Hai-chao;Chen, Qi-xin
椎间盘（IVD）的微环境的特点是基质酸性，缺氧，高渗和营养有限，这是干细胞为基础的再生的主要障碍。我们最近的工作表明，髓核间充质干细胞（NPMSCs）的优势，传统来源的细胞治疗在体外培养样缺氧和高渗条件下。在这里，我们研究的活力，增殖和基质代谢的NPMSCs相比，脂肪组织来源的间充质干细胞（ADMSCs）在体外IVD样酸性条件下。将来自Sprague-Dawley大鼠的ADMSC和NPMSC在代表标准条件（pH 7.4）和正常、轻度退化和严重退化IVD（分别为pH 7.1、6.8和6.5）的四种不同pH水平下培养。通过膜联蛋白-V-异硫氰酸荧光素/碘化丙啶染色检查细胞活力。使用细胞计数试剂盒细胞增殖测定来测量细胞增殖。采用RT-PCR和Western blotting方法检测各组细胞中聚集蛋白聚糖、Ⅰ型胶原、Ⅱ型胶原、基质金属蛋白酶-2（MMP-2）、血小板反应蛋白基序的去整合素和金属蛋白酶-4（ADAMTS 4）和金属蛋白酶组织抑制剂-3（TIMP-3）的mRNA和蛋白水平的表达。在这两种细胞类型中，酸性pH抑制细胞活力和增殖，下调聚集蛋白聚糖，胶原-I，胶原-II和TIMP-3的表达，并上调MMP-2和ADAMTS 4的表达。与ADMSCs相比，NPMSCs在活力和增殖方面受到的抑制明显较少;它们表达的聚集蛋白聚糖和胶原蛋白-II水平明显较高，MMP-2和ADAMTS 4水平较低。因此，酸性环境是ADMSC或NPMSC再生IVD的主要障碍。NPMSCs对酸性pH的抑制不太敏感，可能是基于细胞的IVD再生的有希望的候选者。(C)2015 S. Karger AG，巴塞尔
The microenvironment of the intervertebral disc (IVD) is characterized by matrix acidity, hypoxia, hyperosmolarity and limited nutrition, which are major obstacles to stem cell-based regeneration. Our recent work showed that nucleus pulposus mesenchymal stem cells (NPMSCs) had advantages over traditional sources of cell therapy under IVD-like hypoxic and hyperosmotic conditions. Here, we examined the viability, proliferation and matrix metabolism of NPMSCs compared with adipose tissue-derived mesenchymal stem cells (ADMSCs) under IVD-like acidic conditions in vitro. ADMSCs and NPMSCs from Sprague-Dawley rats were cultured at four different pH levels representing the standard condition (pH 7.4) and the normal, mildly degenerated and severely degenerated IVD (pH 7.1, 6.8 and 6.5, respectively). Cell viability was examined by annexin-V-fluorescein isothiocyanate/propidium iodide staining. Cell proliferation was measured using a cell counting kit cell proliferation assay. The expression of aggrecan, collagen-I, collagen-II, matrix metalloproteinase-2 (MMP-2), a disintegrin and metal-loproteinase with thrombospondin motifs-4 (ADAMTS4) and the tissue inhibitor of metalloproteinase-3 (TIMP-3) was measured at mRNA and protein levels by RT-PCR and Western blotting. In both cell types, acidic pH inhibited cell viability and proliferation, downregulated the expression of aggrecan, collagen-I, collagen-II and TIMP-3, and upregulated the expression of MMP-2 and ADAMTS4. Compared with ADMSCs, NPMSCs were significantly less inhibited in viability and proliferation; they expressed significantly higher levels of aggrecan and collagen-II, and lower levels of MMP-2 and ADAMTS4. Thus, an acidic environment is a major obstacle for IVD regeneration by ADMSCs or NPMSCs. NPMSCs appeared less sensitive to inhibition by acidic pH and might be promising candidates for cell-based IVD regeneration. (C) 2015 S. Karger AG, Basel