Efficiency of Gene Transfection with Adeno-associated Viral Vector or Lentiviral Vector in Bone Marrow Derived Mesenchymal Stem Cells / 中国康复理论与实践

ABSTRACT

Objective To compare 2 kinds of commonly used viral vectors, adeno-associated viral (AAV) vector and lentiviral (LV) vec-tor 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 medi-ated gene transfection, the LV particles were used directly. The transfection efficiency was estimated withβ-gal staining and green fluores-cent 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 prolifer-ating. Some MSCs expressed Nestin. The gene transfection efficiency mediated with AAV vector was 49.1%, and it was 91.4%with LV vec-tor (P<0.01). Conclusion The LV vector is more efficient on gene transfection than AAV vector.