Production and Concentration of Lentivirus for Transduction of Primary Human T Cells.
Lentiviral vectors have emerged as efficient tools for investigating T cell biology through their ability to efficiently deliver transgene expression into both dividing and nondividing cells. Such lentiviral vectors have the potential to infect a wide variety of cell types. However, despite this advantage, the ability to transduce primary human T cells remains challenging and methods to achieve efficient gene transfer are often time consuming and expensive. We describe a method for generating lentivirus that is simple to perform and does not require the purchase of non-standard equipment to transduce primary human T cells. Therefore, we provide an optimized protocol that is easy to implement and allow transduction with high efficiency and reproducibility.
Institute of Immunity and Transplantation, University College London Division of Infection and Immunity, London, NW3 2PF, UK.
Alan
A
Kennedy
Kennedy Institute of Rheumatology, University of Oxford, Roosevelt Drive, Headington, Oxford, OX3 7FY, UK. adam.cribbs@dpag.ox.ac.uk.|Botnar Research Centre, Nuffield Orthopaedic Centre, Windmill Road, Oxford, OX3 7LD, UK. adam.cribbs@dpag.ox.ac.uk.|Department of Physiology, Anatomy and Genetics, MRC Functional Genomics Unit, Computational Genomics and Training Centre (CGAT), Parks Road, Oxford, OX1 3PR, UK. adam.cribbs@dpag.ox.ac.uk.
Adam P
AP
Cribbs
D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D061251:Primary Cell Culture; D013601:T-Lymphocytes / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection / Q000379:methods*; D019076:Transgenes
CD4+ T cells; CD45RA+; Human T cell; Lentiviral vector; Lentivirus; Primary; Titer