Retronectin-assisted retroviral transduction of primary human T lymphocytes under good manufacturing practice conditions: tissue culture bag critically determines cell yield.
BACKGROUND
For 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-foldtrade mark cell culture bags.


METHODS
In 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.


RESULTS
In static transductions, but not in spinoculation, the DC-generation bags and CultiLife Spin bags performed as well as Lifecell X-foldtrade mark 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-foldtrade mark cell culture bags in terms of cell yield.


CONCLUSIONS
Adjusted 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.
Laboratory of Clinical and Tumor Immunology, Erasmus MC-Daniel den Hoed Cancer Center, Rotterdam, The Netherlands. c.lamers@erasmusmc.nl
C H J
CH
Lamers
P
P
van Elzakker
S C L
SC
van Steenbergen
S
S
Sleijfer
R
R
Debets
J W
JW
Gratama
D000818:Animals; D002292:Carcinoma, Renal Cell / Q000276:immunology / Q000628:therapy; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D008422:Materials Testing; D012190:Retroviridae / Q000235:genetics / Q000378:metabolism*; D013601:T-Lymphocytes / Q000166:cytology / Q000502:physiology*; D014161:Transduction, Genetic / Q000379:methods*