Application of a continuous bioreactor cascade to study the effect of linoleic acid on hybridoma cell physiology

Abstract

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 microM linoleic acid. At the end of the experiment with 50 microM linoleic acid, the concentration of linoleic acid is increased stepwise to determine the cytotoxic level. For rapid proliferating cells exposed to 25 and 50 microM stimulation of growth was observed. At 50 microM 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 microM and arrest of cell proliferation in the G(2)/M phase at 50 microM. For both, rapid proliferating cells and stressed apoptotic cells, complete growth inhibition occurred at 85 microM, 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 microM; IC(100) = 400 microM). 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.

Bioreactor Process Parameter Screening Utilizing a Plackett-Burman Design for a Model Monoclonal Antibody

“…3). This finding is consistent with prior published observations including dose‐dependent fatty acid supplementation having neutral or positive effects at lower concentrations (<25 mmol/L), decreased glutamine metabolism of hybridoma cells resulting in slowed growth kinetics, and increased shear‐induced apoptosis occurring at higher fatty acid supplementation concentrations (>50 mmol/L) …”

“…This finding is consistent with prior published observations including dosedependent fatty acid supplementation having neutral or pos- itive effects at lower concentrations (<25 mmol/L), decreased glutamine metabolism of hybridoma cells resulting in slowed growth kinetics, and increased shear-induced apoptosis occurring at higher fatty acid supplementation concentrations (>50 mmol/L). 37,38 A significant effect was seen with hydrocortisone supplementation, which resulted in an increase in the total cell count near the end of culture, that is, day 4 TC/mL, and the maximum TC/mL measured during culture was significantly increased by the drip-feed strategy ( Table 3). The apparent increases in TC/mL were not seen in corresponding measures of VC/mL, and likely reflect accumulation of dead cells.…”

Impact of bioaccumulated selenium on nutraceutical properties and volatile compounds in submerged fermented Pleurotus eryngii mycelia

“…And 9,12‐octadecadienoic acid/Linoleic acid (MW‐280) was found as a major compound to be present in both untreated sample (peak area‐53.47%) and the treated sample (peak area – 32.75%). Linoleic acid being a poly‐unsaturated fatty acid known for its medicinal bioactivities like anticancer, anti‐inflammatory, antihypercholesterolemic, hepatoprotective, antiarthritic, and anti‐coronary (Jordão Candido et al, 2019; Kisztelinski et al, 2006). N‐Hexadecanoic acid content which possesses antihypercholesterolemic and antioxidative properties has also increased significantly in treated samples.…”

Chemical composition, antioxidant and antitumor activities of sub-fractions of wild and cultivated Pleurotus ferulae ethanol extracts

“…Linoleic acid and its methyl esters have many beneficial effects in the prevention of atherosclerosis, cancer, hypertension and improvement of immune function (Whelan, 2008; Bhattacharya et al, 2006). It has been reported that linoleic acid or conjugated linoleic acid can inhibit the proliferation of hybridoma cells or human breast cancer MCF74 cells through induction of apoptosis or activation of p53, respectively (Albright et al, 2005; Kisztelinski et al, 2006). These studies suggest that methyl linoleate may be associated with the antitumor effects of Pe-W and Pe-C.…”