Validation of an in vitro nutrition model using an enteral formula in aged neutrophils.

The aim of the study is to validate a cell culture model appropriate for assessing the effects of standard nutritional formulas on neutrophil functionality in vitro. The model consists of aged cells exposed to a commercial nutritional formula containing solely LCT as lipid component. Preliminary experiments determined dosage of formula and culture interval. Neutrophils were isolated from a pool of whole blood in healthy volunteers (18-55 years old) and cultured with and without addition of a commercial enteral diet with 3.5% lipids (equivalent to 0.04, 0.08, 0.2 and 0.4 mM of intraassay LCT) for 18, 42 or 76 hours. Based on cell viability results, doses of 0.2 and 0.4 mM LCT and culture time of 18 hours were established for subsequent experiments. Neutrophil functionality was evaluated by phagocytosis (NBT test), MDA production (lipoperoxidation index) and DNA fragmentation. Optic microscopy showed higher percentages of pre-apoptotic cells and a significant increase in DNA fragmentation as compared to controls only with an LCT concentration of 0.4 mM (p < 0.05). Interestingly, cell cultures with both 0.2 and 0.4 mM of added LCT showed significant decreases in malonyldialdehyde (MDA) release as a lipoperoxidation marker. This nutrition model of neutrophils and in vitro complete enteral commercial diet is relatively simply to execute and can be applied to different pathological conditions in which the aim is to study changes in neutrophil functionality.