Culture of human tumor infiltrating lymphocytes in hollow fiber bioreactors.
Human tumor infiltrating lymphocytes (TIL) from metastatic melanoma of six patients were grown using a new hollow fiber bioreactor system. After inoculating 0.35-10 X 10(8) TIL into the extra-fiber space (EFS), each Cellmax bioreactor was perfused with AIM-V medium, supplemented with rIL-2. The cells subsequently expanded 124-1170-fold to yield 1.5-5.4 X 10(10) TIL over a 14-32 day period. TIL were flushed from the EFS using 200 ml medium and possessed an average viability = 91%. The phenotype and the autologous tumor cell lytic capacity of these TIL were similar to those of TIL grown in the currently used gas-permeable culture bags. Tissue culture media use averaged 4.3 liters/10(10) TIL harvested. The TIL of one patient were re-expanded twice from cells remaining within the same bioreactor after harvest suggesting that one bioreactor cartridge could be used for repetitive, periodic studies. An estimated 80% decrease in technical time expended and in incubator space requirements were realized using this methodology. Cell culture on hollow fibers appears to be a useful method for producing large quantities of primary human lymphocytes for experimental, and perhaps, therapeutic needs.
Knazek
R A
RA
Cellco Advanced Bioreactors, Inc., Kensington, MD 20895.
Wu
Y W
YW
Aebersold
P M
PM
Rosenberg
S A
SA
D002478:Cells, Cultured; D003470:Culture Media; D003602:Cytotoxicity, Immunologic; D006801:Humans; D008545:Melanoma / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
D003470:Culture Media