Efficiency of Gene Transfection with Adeno- associated Viral Vector or Lentiviral Vector in Bone Marrow Derived Mesenchymal Stem Cells
10.3969/j.issn.1006-9771.2014.12.005
- VernacularTitle: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
- Author:
Yujin SU
;
Yumei ZHAO
;
Yi GU
- Publication Type:Journal Article
- Keywords:
gene transfection, adeno-associated viral vector, lentiviral vector, mesenchymal stem cells
- From:
Chinese Journal of Rehabilitation Theory and Practice
2014;20(12):1117-1121
- CountryChina
- Language:Chinese
-
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.
