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Application of a continuous bioreactor cascade to study the effect of linoleic acid on hybridoma cell physiology
Author(s) -
Kisztelinski Dominik,
Alink Gerrit M.,
Rietjens Ivonne M.C.M.,
Bielecki Stanislaw,
Tramper Johannes,
Martens Dirk E.
Publication year - 2006
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20897
Subject(s) - linoleic acid , bioreactor , apoptosis , cell growth , cell , cell culture , biology , biochemistry , chemistry , microbiology and biotechnology , fatty acid , botany , genetics
The aim of the present study is to demonstrate the use of controlled bioreactors for toxicological studies. As a model system the effect of linoleic acid on hybridoma cells is studied in two well‐controlled continuously operated bioreactors placed in series. In the first reactor the effect on rapid proliferating cells can be studied, while in the second reactor a special steady state is created, which allows studying the effect on apoptotic cells. Experiments are done at 0, 25, and 50 µM linoleic acid. At the end of the experiment with 50 µM linoleic acid, the concentration of linoleic acid is increased stepwise to determine the cytotoxic level. For rapid proliferating cells exposed to 25 and 50 µM stimulation of growth was observed. At 50 µM there was at the same time an increase in cell death through apoptosis. For stressed apoptotic cells linoleic acid caused partial growth inhibition at 25 and 50 µM and arrest of cell proliferation in the G 2 /M phase at 50 µM. For both, rapid proliferating cells and stressed apoptotic cells, complete growth inhibition occurred at 85 µM, with cells being arrested in the G 2 /M phase and dying mainly through necrosis. Cells in the bioreactor system appeared to be more sensitive towards linoleic acid than cells grown in multi‐well plates. (IC 50 = 300 µM; IC 100 = 400 µM). Altogether the results of the present study reveal that the biostat experiments allow detailed analysis of the effect of a bioactive ingredient on cell physiology and behavior. © 2006 Wiley Periodicals, Inc.