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国家自然科学基金(30750010)

作品数:2 被引量:5H指数:2
相关作者:胡勇王庆德赵继军潘晓华杨述华更多>>
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低氧抑制人骨髓间充质干细胞向成骨细胞分化被引量:2
2008年
目的探讨低氧对人间充质干细胞(hMSCs)分化为成骨细胞的影响,为骨组织工程学提供实验依据。方法根据培养氧浓度及培养液类型随机分为4组:正常氧组(n)(20%O2+DMEM-LG),低氧组(h)(1%O2+DMEM-LG),正常氧成骨诱导组(nos)(20%O2+条件培养液)及低氧成骨诱导组(hos)(1%O2+条件培养液)。观察细胞形态变化,检测碱性磷酸酶(ALP)活性,定量分析ALP、Ⅰ型胶原蛋白(COL1A2)、骨钙素(OC)及骨粘连蛋白(BSP)mRNA表达情况,钙结节使用茜素红染色。结果与nos组相比,n、h及hos组hMSCs生长形态改变不明显,并且不受条件培养液的影响。hos组hMSCs的ALP活性逐渐增高,但明显低于nos组,两者差异有统计学意义(P<0.01);培养4周后,与其它组相比,nos组的hMSCs可见到明显的染成红色的钙结节及钙盐沉积。定量RT-PCR检测,nos组hMSCs的ALP、OC、COL1A2及BSP mRNA表达量从7d时开始明显增加,ALP14d后达到峰值,随后活性开始降低,OC、COL1A2及BSP mRNA继续增高,hos组ALP及BSP mRNA14d后开始增加,但明显低于nos组(均P<0.05)。结论低氧环境在体外抑制hMSCs成骨细胞分化,延迟hMSCs成骨。
王庆德杨述华杨操胡勇赵继军潘晓华
关键词:间充质干细胞低氧成骨细胞细胞培养
Co-culture of Mesenchymal Stem Cells with Umbilical Vein Endothelial Cells under Hypoxic Condition被引量:3
2012年
By co-culturing humm mesenchymal stem cells (hMSCs) and human umbilical rein endothelial cells (HUVECs) under hypoxia and creating a microenvironment similar to that of transplanted hMSCs for the treatment of avascular ni ANFH, the effect of hMSCs on survival, apoptosis, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) under the hypoxic condition were investigated in vitro. hMSCs and HUVECs were cultured and identified in vitro. Three kinds of conditioned media, CdM-CdMNOR, CdM-CdMHYP and HUVEC-CdMHYP were prepared. HUVECs were cultured with these conditioned media under hypoxia. The survival rate, apoptosis rate, migration and angiogenesis of HUVECs were respectively detected by CCK-8, flow cytometry, Transwell and tube formation assay. The content of SDF-1α, VEGF and IL-6 in CdM was determined by ELISA. Our results showed that hMSCs and HUVECs were cultured and identified successfully. Compared with MSC-CdMNOR and HUVEC-CdMHYP groups, the survival rate, migra-tion and angiogenesis of HUVECs in MSC-CdMHYP group were significantly increased while the apoptosis rate was declined (P<0.05). Moreover, the expression of SDF-1α, VEGF and IL-6 in MSC-CdMHYP group was up-regulated. Under hypoxia, the apoptosis of HUVECs was inhibited while survival, migration and angiogenesis were improved by co-culture of hMSCs and HUVECs. The underlying mechanism may be that hMSCs could secrete a number of cytokines and improve niche, which might be helpful in the treatment of femoral head necrosis.
张波杨述华张宇坤孙志博许伟华叶树楠
关键词:HYPOXIACO-CULTURE
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