基础医学与临床 ›› 2016, Vol. 36 ›› Issue (7): 924-928.

• 研究论文 • 上一篇    下一篇

复合基因载体可安全高效转导大鼠骨髓间充质干细胞

万莉1,姚醒蕾1,张添源2,高建青2,韩钦3,赵春华4   

  1. 1. 中国医学科学院基础医学研究所
    2. 浙江大学 药学院药物制剂研究所
    3. 中国医学科学院基础医学研究所组织工程中心
    4. 中国医学科学院基础研究所组织工程中心
  • 收稿日期:2016-04-08 修回日期:2016-05-19 出版日期:2016-07-05 发布日期:2016-06-22
  • 通讯作者: 赵春华 E-mail:chunhuaz@public.tpt.tj.cn.com
  • 基金资助:
    国家自然科学基金;国家重大科学研究计划

Combined gene vector can transduce rat mesenchymal stem cells efficiently and safely

  • Received:2016-04-08 Revised:2016-05-19 Online:2016-07-05 Published:2016-06-22
  • Contact: Chun-hua ZHAO, E-mail:chunhuaz@public.tpt.tj.cn.com
  • Supported by:
    ;The National key scientific research program

摘要: 目的 探讨精胺普鲁兰多糖多聚体(SP)与腺病毒载体(Adv)形成的复合基因载体(SP-Adv)对大鼠骨髓间充质干细胞(MSCs)的转导效率及其安全性。方法 SP与腺病毒载体(Adv)室温混合孵育15 min形成SP-Adv;透射电镜观察SP-Adv的物理化学性质;双萤光素酶报告系统检测SP-Adv在MSCs中的转导效率;MTT实验评价SP-Adv的细胞毒性;碱性磷酸酶染色和油红O染色检测SP-Adv对MSCs成骨及成脂分化能力的影响。结果 SP-Adv粒径变大且原病毒纤突消失被水合层代替;SP-Adv对MSCs的基因转导效率显著高于未包裹的Adv(P<0.001),且SP-Adv对MSCs无明显的细胞毒性;MSCs的成骨细胞和成脂细胞分化实验证实SP-Adv对MSCs的分化能力无影响。结论 SP-Adv可以对MSCs进行高效的基因转导,且证实该转导在细胞水平具有安全性。

关键词: 关键词:间充质干细胞, 复合基因载体, 基因转导

Abstract: Objective To explore the transgene efficiency and safety of combined vector of spermine-pullulan polymer (SP) and adenovirus vector (Adv) on rat bone marrow derived MSCs (MSCs). Methods To combine vectors, Adv was incubated with the SP at room temperature for 15 min to form SP-Adv, and the morphology was observed under a transmission electron microscope. Luciferase activity reflecting the transduction efficiency was determined by using a luciferase assay system, and cell viability was determined using an MTT assay system. The osteogenic differentiation was detected by alkaline-phosphatase staining, and adipogenic differentiation was detected by Oil-Red-O staining. Results We observed that Adv was coated with SP polymer and the size of SP-Adv was bigger than uncoated Adv. SP-Adv showed significantly higher transgene expression than uncoated Adv and did not affect cell viability of MSCs. SP-Advs did not impair the osteogenic nor adipogenic differentiation of MSCs. Conclusion SP-Adv can transduce mesenchymal stem cells efficiently, and their safety was demonstrated in cell level.

Key words: Keyword: Mesenchymal stem cells, Combined gene vector, Gene transduction

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