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Molecular Biotechnology

, 43:250 | Cite as

Gene Delivery Efficiency in Bone Marrow-derived Dendritic Cells: Comparison of Four Methods and Optimization for Lentivirus Transduction

  • Gong-Bo Li
  • Guang-Xiu Lu
Research

Abstract

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.

Keywords

Dendritic cell Lentivirus Gene delivery 

Abbreviations

BMDC

Bone marrow-derived dendritic cell

GM-CSF

Granulocyte–macrophage colony-stimulating factor

rLV

Recombinant lentivirus

rAAV2

Recombinant adeno-associated virus type 2

MOI

Multiplicity of infection

TU

Transduction unit

APC

Antigen presenting cell

Notes

Acknowledgment

This study was supported by The Hi-Tech Research and Development of China, Grant No. 2006AA02A102.

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Copyright information

© Humana Press 2009

Authors and Affiliations

  1. 1.Institute of Human Reproduction and Stem Cell EngineeringCentral South University, National Engineering and Research Center of Human Stem CellChangshaChina

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