Simulated microgravity inhibits the proliferation and osteogenesis of rat bone marrow mesenchymal stem cells
Simulated microgravity inhibits the proliferation and osteogenesis of rat bone marrow mesenchymal stem cells
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DOI：
10.1111/j.1365-2184.2007.00461.x
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发表时间：
2007-10-01
影响因子：
8.5
通讯作者：
Li, Y. H.
中科院分区：
文献类型：
--
作者：
Dai, Z. Q.;Wang, R.;Li, Y. H.
目的：已知微重力会影响骨髓间充质干细胞（BMSC）的分化。然而，最近报道了一些关于微重力对骨髓间充质干细胞增殖的影响的有争议的发现。因此，我们研究了模拟微重力对大鼠 BMSC (rBMSC) 增殖及其成骨潜力的影响。材料和方法：使用我们建立的基于红细胞裂解的有效方法从骨髓中分离出rBMSCs，通过其表面标志物及其在正常条件下的增殖特性来鉴定。然后，将它们在有或没有生长因子的回转器中模拟微重力以及在成骨培养基中培养。随后，分别使用亚甲蓝染色和流式细胞术评估增殖和细胞周期参数；使用蛋白质印迹和微阵列分析测定基因表达。结果：模拟微重力抑制 rBMSC 群体生长，细胞被停滞在细胞周期的 G(0)/G(1) 期。生长因子，如胰岛素样生长因子-I、表皮生长因子和碱性成纤维细胞生长因子，在正常重力下可显着刺激rBMSC增殖，但在模拟微重力下效果轻微。旋转培养 3 天后，Akt 和细胞外信号相关激酶 1/2 磷酸化水平以及核心结合因子 α 1 的表达下降。微阵列和基因本体分析进一步证实rBMSC增殖和成骨作用在模拟微重力下下降。结论：上述数据表明模拟微重力抑制rBMSCs的群体生长及其向成骨细胞的分化。这些变化可能是太空飞行期间注意到的一些生理变化的原因。
Objectives: Microgravity is known to affect the differentiation of bone marrow mesenchymal stem cells (BMSCs). However, a few controversial findings have recently been reported with respect to the effects of microgravity on BMSC proliferation. Thus, we investigated the effects of simulated microgravity on rat BMSC (rBMSC) proliferation and their osteogeneic potential. Materials and methods: rBMSCs isolated from marrow using our established effective method, based on erythrocyte lysis, were identified by their surface markers and their proliferation characteristics under normal conditions. Then, they were cultured in a clinostat to simulate microgravity, with or without growth factors, and in osteogenic medium. Subsequently, proliferation and cell cycle parameters were assessed using methylene blue staining and flow cytometry, respectively; gene expression was determined using Western blotting and microarray analysis. Results: Simulated microgravity inhibited population growth of the rBMSCs, cells being arrested in the G(0)/G(1) phase of cell cycle. Growth factors, such as insulin-like growth factor-I, epidermal growth factor and basic fibroblastic growth factor, markedly stimulated rBMSC proliferation in normal gravity, but had only a slight effect in simulated microgravity. Akt and extracellular signal-related kinase 1/2 phosphorylation levels and the expression of core-binding factor alpha 1 decreased after 3 days of clinorotation culture. Microarray and gene ontology analyses further confirmed that rBMSC proliferation and osteogenesis decreased under simulated microgravity. Conclusions: The above data suggest that simulated microgravity inhibits population growth of rBMSCs and their differentiation towards osteoblasts. These changes may be responsible for some of the physiological changes noted during spaceflight.