《中国康复理论与实践》

《中国康复理论与实践》 ›› 2014, Vol. 20 ›› Issue (12): 1117-1121.

• 础基研究 • 上一篇    下一篇

腺相关病毒及慢病毒载体对骨髓间充质干细胞基因转染效率的比较

苏玉金，赵育梅，顾漪
   

  1. 北京市神经外科研究所，首都医科大学附属北京天坛医院，北京市100050。
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2014-12-25 发布日期:2014-12-25

Efficiency of Gene Transfection with Adeno- associated Viral Vector or Lentiviral Vector in Bone Marrow Derived Mesenchymal Stem Cells

SU Yu- jin, ZHAO Yu- mei, GU Yi.
  

  1. Beijing Neurosurgical Institute, Tiantan Hospital, Capital Medical University, Beijing 100050, China
  • Received:1900-01-01 Revised:1900-01-01 Published:2014-12-25 Online:2014-12-25

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被引次数

10

摘要： 目的比较腺相关病毒(AAV)载体和慢病毒(LV)载体在间充质干细胞(MSCs)基因转染中的效率。方法密度梯度离心法分离培养MSCs，HE 染色、Nestin 免疫荧光染色鉴定，BrdU 标记观察增殖情况。分别包装AAV 与LV 假病毒颗粒，并感染MSCs，通过β-gal 染色和绿色荧光蛋白检测，分别计算两者的转染效率。结果MSCs成功从骨髓分离。AAV载体介导的基因转染率为49.1%，LV载体的转染率为91.4% (P<0.01)。结论LV载体介导的基因转染方法转染效率更高。

关键词: 基因转染, 腺相关病毒载体, 慢病毒载体, 骨髓间充质干细胞

Abstract: Objective To compare 2 kinds of commonly used viral vectors, adeno-associated viral (AAV) vector and lentiviral (LV) vector in the gene transfection for bone marrow derived mesenchymal stem cells (MSCs). Methods MSCs were isolated with density gradient (lymphocytes seperation) and identified with HE staining and immunocytochemistory staining for Nestin. The proliferation of BMSCs was detected with BrdU labeling. AAV mediated gene transfection was carried out through recombinant AAV-LacZ viral particles. For LV mediated gene transfection, the LV particles were used directly. The transfection efficiency was estimated with β-gal staining and green fluorescent protein under the fluorescent microscope respectively. Results MSCs was successfully isolated from the bone marrow. HE staining showed that MSCs was with big nucleus, 1-3 nucleoli, and high nucleocytoplasmic ratio. BrdU labeling suggested that MSCs were proliferating. Some MSCs expressed Nestin. The gene transfection efficiency mediated with AAV vector was 49.1%, and it was 91.4% with LV vector (P<0.01). Conclusion The LV vector is more efficient on gene transfection than AAV vector.

Key words: gene transfection, adeno-associated viral vector, lentiviral vector, mesenchymal stem cells