Natural Killer Cell Membrane Isolation with Minimal Nuclear, Mitochondrial, and Cytosol Contamination

Cell membrane isolation is essential for diverse biological investigations, ranging from fundamental research to advanced therapeutic applications. This study compared two methods-differential centrifugation and discontinuous sucrose density gradient ultracentrifugation-for isolating cell membranes from the human natural killer (NK) cell line (KHYG-1). The aim was to identify the method that minimizes contamination from nuclear, mitochondrial, and cytosolic components. Differential centrifugation yielded approximately 8 mg of cell membrane, whereas sucrose density gradient ultracentrifugation produced about 5 mg. Despite the lower yield, the latter method exhibited superior performance due to significantly reduced contamination. This protocol is adaptable to various cell types, offering a reliable approach for producing cell membrane-coated mimics for therapeutic use. The increasing demand for isolated cell membranes in biomedical applications highlights the importance of optimizing isolation techniques.