We have previously designed and validated a GMP protocol for retroviral gene transduction
and expansion of human T-cells. Here, we have revisited the choice for optimal medium
(composition) and tested different and newly available media for their effects on
yield as well as phenotypical and functional properties of receptor-engineered T-cells.
GMP-defined (serum-free) media that were tested included AIMV and Optimize (Invitrogen),
Xvivo15 (Lonza), CellGro (CellGenix) and TexMACS(Miltenyi). We tested these media
with or without supplementation of 2% plasma. In addition, we tested 3 blended media
(i) AIMV (20%)+RPMI(80%)+2% plasma (EMC standard); (ii) AIMV(50%)+RPMI(50%)+5% human
serum; and (iii) Xvivo15(20%)+RPMI(80%)+2% plasma. First experiments used CAR-engineered
T-cells.The non-supplemented media performed the least with respect to T-cell yield,
transduction efficiency and function of CAR T-cells. The best performing media with
respect to these parameters included 3 plasma-supplemented media (AIMV, Xvivo15 and
TexMACS) and the 3 blended media, including the EMC standard. Five of these media,
supplemented with IL15+IL21, were re-tested in parallel to generate MC2/A2 TCR T-cells.
In terms of T-cell yield, transduction and function, AIMV+2% plasma performed the
best, whereas Xvivo+2% plasma performed the least. Notably, with regard to the preservation
of early T-cell differentiation this sequence was reverse. Taken together, this study
shows the impact of culture media on yield and phenotypical and functional properties
of human T-cells. Importantly, the medium resulting in highest T-cell yield led to
more differentiated T-cells, and vice versa. This study provides important information
with respect to medium selection to gene-engineer and process T-cells for clinical
studies.
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© 2014 Published by Elsevier Inc.