Abstract: Objective To compare the three different methods for direct transfection of small exogenous nicotinamide adenine dinucleotide phosphate(NADPH) into cells by a non-metabolic pathway. Methods Three different transfection reagents (X-tremeGENE^TM HP DNA, Lipofectamine^TM RNAiMAX and Lipofectamine^TM 2000) were used to find the transfection efficiency of NADPH into human osteosarcoma cell line U2OS and mouse embryonic fibroblast cell line 3T3L1. Their effects on adipocyte differentiation were compared by Oil Red O staining. Results Transfection of NADPH with X-tremeGENE HP DNA transfection reagent effectively increased intracellular NADPH level(P＜0.001). With the increase of NADPH transfection concentration (10 μmol/L NADPH and 10 μL transfection reagent), the level of NADPH in cells increased in a dose-dependent manner. In addition, three transfection reagents were used to transfect NADPH in 3T3L1 preadipocytes. Only cells transfected with NADPH using X-tremeGENE HP DNA transfection reagent contained more lipids than control cells (P＜0.001). Conclusions Transfection of NADPH with X-tremeGENE HP DNA transfection reagent effectively increases the level of NADPH in cells.

Key words: nicotinamide adenine dinucleotide phosphate(NADPH), transfection, cell culture, adipocyte differentiation