腺病毒构建过表达睫状神经营养因子对骨髓间充质干细胞的影响
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国家自然科学基金资助项目(No.81770948); 福建省科技创新领军人才基金资助项目(No.2016B011)


Effects of ciliary neurotrophic factor over-expression by adenovirus on bone marrow mesenchymal stem cells
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National Natural Science Foundation of China(No.81770948); Fujian Provincial Science and Technology Innovation Leadership Talent Foundation(No.2016B011)

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    摘要:

    目的:构建过表达睫状神经营养因子(CNTF)的腺病毒,利用腺病毒载体转染骨髓间充质干细胞,使其在体外持续高效表达CNTF,为视网膜病变的体外研究提供新途径。

    方法:构建空载GFP-腺病毒及CNTF-腺病毒,取第3代BMSCs用于转染实验,分为3组:空白对照组(未转染腺病毒组)、阴性对照组(GFP-腺病毒转染组)和实验组(CNTF-腺病毒转染组)。ELISA测转染后1、2、3d各组上清液的CNTF蛋白分泌量。

    结果:成功构建空载GFP-腺病毒及CNTF-腺病毒, 实验组的CNTF表达量明显高于空白对照组和阴性对照组,差异具有统计学意义(P<0.05)。

    结论:利用腺病毒载体可使BMSCs在体外持续高效表达CNTF。

    Abstract:

    AIM: To establish the model of bone marrow mesenchymal stem cells(BMSCs)over-expressing ciliary neurotrophic factor(CNTF)by applying adenovirus. It may provide novel strategies for in-vitro investigations of retinopathies.

    METHODS: GFP-adenovirus and CNTF-adenovirus were synthesized then used to transfect BMSCs of passage 3. Blank control group(without adenovirus transfected group), negative control group(GFP-adenovirus transfected group), and experimental group(CNTF-adenovirus transfected group)were included in this study. On the 1, 2, 3d post-transfected, ELISA assay was applied to examine CNTF protein-secretion in the supernate.

    RESULTS: GFP-adenovirus and CNTF-adenovirus models were successfully established. The CNTF protein levels in the supernate were higher in experimental group than those in the blank control group and negative control group(P<0.05).

    CONCLUSION: CNTF-modified BMSCs by adenovirus could efficiently secrete CNTF protein in-vitro.

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林雯,徐国兴.腺病毒构建过表达睫状神经营养因子对骨髓间充质干细胞的影响.国际眼科杂志, 2020,20(12):2039-2043.

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  • 收稿日期:2020-05-24
  • 最后修改日期:2020-11-11
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  • 在线发布日期: 2020-11-19
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