Retronectin®-assisted retroviral transduction of primary human T lymphocytes under good manufacturing practice conditions: tissue culture bag critically determines cell yield

BackgroundFor our clinical immunogene therapy study for the treatment of renal cell carcinoma (RCC) patients, we had developed a protocol for gene transduction and expansion of human T cells in compliance with good manufacturing practice (GMP) criteria. Critical to our successful clinical-scale transductions of patient T cells was the use of Retronectin® in combination with Lifecell® X-fold™ cell culture bags.MethodsIn our current study, we evaluated two alternative types of bags for the Retronectin®-mediated retroviral transduction of human T cells: the Miltenyi DC-generation bag and the Takara CultiLife Spin bag.ResultsIn static transductions, but not in spinoculation, the DC-generation bags and CultiLife Spin bags performed as well as Lifecell® X-fold™ bags in Retronectin®-assisted retroviral transduction of human T cells with respect to transduction efficiency, lymphocyte subset composition and lymphocyte function. However, both types of bags performed less well than Lifecell® X-fold™ cell culture bags in terms of cell yield.DiscussionAdjusted numbers of cells at the start of transduction should be used when using the Miltenyi or Takara bags in order to compensate for the lower cell yield following transduction.