不同诱导条件对人骨髓基质干细胞向内皮分化的影响
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国家自然科学基金资助项目(30400517和30270568)


Plasticity of Bone Marrow Stromal Stem Cells along Endothelial Lineage under Different Culture Condition
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    目的探讨不同诱导条件对人骨髓基质干细胞向内皮分化的影响。方法采用密度梯度离心法分离培养人骨髓基质干细胞,用荧光激活细胞分选法分析骨髓基质干细胞CD34、CD105和CD166的表达率;用免疫荧光细胞化学法观察骨髓基质干细胞在细胞因子(50μgL血管内皮细胞生长因子、5μgL碱性成纤维细胞生长因子、100mgL内皮细胞生长补充因子)、内皮条件培养基或与成熟兔主动脉内皮共培养5天时vWF或和Flk1蛋白的表达。结果分离培养的骨髓基质干细胞CD34表达率为4.16%±0.16%,与阴性对照(4.06%±0.23%)相比无统计学差异,CD105表达率为90.20%±2.35%,CD166表达率为82.30%±3.22%,均明显高于阴性对照(p<0.05)。诱导前骨髓基质干细胞不表达vWF和Flk1蛋白,经血管内皮细胞生长因子、碱性成纤维细胞生长因子和内皮细胞生长补充因子等细胞因子联合诱导5天时,33.42%骨髓基质干细胞开始表达vWF蛋白;与成熟内皮共培养5天时,vWF染色仍为阴性,但25.71%骨髓基质干细胞开始表达Flk1;用内皮条件培养基培养5天时,骨髓基质干细胞vWF和Flk1染色均为阴性。结论细胞因子和成熟内皮细胞均能诱导骨髓基质干细胞向内皮分化,而内皮条件培养基不能诱导骨髓基质干细胞向内皮分化。

    Abstract:

    Aim To investigate the plasticity of bone marrow stromal stem cells(MSCs) along endothelial lineage under different culture condition. Methods The human MSCs were isolated by density gradient centrifugation. Fluorescence activated cell sorter (FACS) was used to analyze the phenotypic characteristics of MSCs. Fluorescence Immunocytochemistry was used to observe the vWF and/or Flk-1 protein expression of MSCs after cultured with DMEM medium containing 50 μg/L VEGF, 5 μg/L bFGF, 100 mg/L ECGS or with endothelial cells(ECs) conditioned medium or cocultured with mature rabbit ECs for 5 days. Results FACS analysis indicated that the MSCs did not express CD34 (CD34 expressing cells vs negative control: 4.16%±0.16% vs 4.06%± 0.23 %, n=6, p>0.05), but expressed CD105 and CD166 (CD105 expressing cells vs negative control: 90.20%±2.35% vs 4.06%±0.23%; CD166 expressing cells vs negative control: 82.30%±3.22% vs 4.06%±0.23%, n=6, p<0.05). 33.42% MSCs began to express vWF after cultured in DMEM containing VEGF, bFGF and ECGS. 25.71% MSCs began to express Flk-1, but did not express vWF after cocultured with mature ECs for 5 days. The MSCs did not express Flk-1 or vWF after cultured in ECs conditioned medium. Conclusions Cytokines and mature ECs could induce MSCs to differentiate alone endothelial lineage, while ECs conditioned medium could not.

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武晓静,黄岚,周骐,崔斌,董红梅,周音频.不同诱导条件对人骨髓基质干细胞向内皮分化的影响[J].中国动脉硬化杂志,2005,13(4):393~396.

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  • 收稿日期:2004-10-22
  • 最后修改日期:2005-06-15
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