Effects of intravenous lipid emulsions on Jurkat cells assessed using label-free deformability cytometry

Intravenous lipid emulsions (ILEs) are routinely used in clinical nutritional support and exert immunomodulatory effects, although they remain poorly understood. This is particularly relevant for cell-based immunotherapies, such as CAR-T therapy, where interactions between immune cells and circulating lipids may influence treatment efficacy. We investigated the effects of two commonly used ILEs (SMOFlipid and Omegaven) on Jurkat cells, using deformability cytometry (DC). Beyond assessing cell mechanics, we extended DC with two novel, label-free approaches: deep neural network-based image classification for cell state evaluation and morphological analysis for lipid droplet detection.Our results show that ILE composition strongly affects cytotoxicity. Omegaven, rich in omega-3 polyunsaturated fatty acids, was markedly more toxic than SMOFlipid, which mainly contains omega-6 and omega-9 fatty acids. While cell stiffness remained largely unchanged, DC effectively detected reduced viability and increased lipid droplet accumulation. Seahorse metabolic analysis supported these findings, showing no significant changes in energy metabolism, suggesting that excess lipids were stored rather than metabolized.This study highlights a novel application of DC for high-throughput, label-free characterization of immune cells, providing insights into how ILEs may influence T cell biology and potentially impact the safety and efficacy of immunotherapies.