T cell culture for gammaretroviral transfer.
Gene transfer into mature T cells with gammaretroviral vectors requires prestimulation, as only mitotic cells are susceptible to integration of the gammaretroviral proviral genome. Costimulation via the CD3/ TCR complex and a second costimulatory molecule, such as CD28 was found to better preserve functionality of the T lymphocytes during ex vivo expansion than stimulation with anti-CD3 alone. The protocols described here for prestimulation and transduction of human and murine T cells with gammaretroviral vectors were optimized for high-level gene transfer and maximum yield of functional T lymphocytes.
Newrzela
Sebastian
S
Institute for Biomedical Research, Georg-Speyer-Haus, Frankfurt am Main, Germany.
Gunda
Brandenburg
B
von Laer
Dorothee
D
D000818:Animals; D002478:Cells, Cultured; D003470:Culture Media; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D012190:Retroviridae / Q000235:genetics*; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*
D003470:Culture Media