Gene delivery efficiency in bone marrow-derived dendritic cells: comparison of four methods and optimization for lentivirus transduction.
Many gene delivery methods have been used to transduce or transfect bone marrow-derived dendritic cells (BMDCs) for genetic engineered DC vaccine research. The present study, for the first time, evaluated the efficiencies of four methods (lipofection, DNA electroporation, recombinant adeno-associated virus type 2 (rAAV2) transduction, and recombinant lentivirus (rLV) transduction) using EGFP as a report gene in the same BMDC culture system. Our data demonstrate that rLV transduction is the most effective method; both lipofection and DNA electroporation transfect BMDCs at lower efficiencies; rAAV2 can hardly transduce BMDCs. Furthermore, our results, for the first time, demonstrate that rLV transduction efficiency on BMDCs can be improved significantly by co-centrifugation and repeated transduction.
Li
Gong-Bo
GB
Institute of Human Reproduction and Stem Cell Engineering, Central South University, National Engineering and Research Center of Human Stem Cell, Changsha, 410078, China.
Lu
Guang-Xiu
GX
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology*; D003713:Dendritic Cells / Q000166:cytology / Q000502:physiology*; D005260:Female; D005434:Flow Cytometry; D006583:Hexadimethrine Bromide; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D014162:Transfection / Q000379:methods*
D006583:Hexadimethrine Bromide