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

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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.

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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

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Acknowledgment

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

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  1. Institute of Human Reproduction and Stem Cell Engineering, Central South University, National Engineering and Research Center of Human Stem Cell, 110 Xiangya road, Changsha, 410078, China

    Gong-Bo Li & Guang-Xiu Lu

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  1. Gong-Bo Li
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  2. Guang-Xiu Lu
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Correspondence to Guang-Xiu Lu.

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Li, GB., Lu, GX. Gene Delivery Efficiency in Bone Marrow-derived Dendritic Cells: Comparison of Four Methods and Optimization for Lentivirus Transduction. Mol Biotechnol 43, 250–256 (2009). https://doi.org/10.1007/s12033-009-9197-1

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  • DOI: https://doi.org/10.1007/s12033-009-9197-1

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