Cell membrane rupture: a novel test reveals significant variations among different brands of tissue culture flasks

OBJECTIVES: Loss of cytoplasmic molecules including protein controls, due to cell membrane rupture can cause errors and irreproducibility in research data. Previous results have shown that during the washing of a monolayer of cells with a balanced salt solution, the fluid force causes cell membrane rupture on some areas of the flasks/dishes. This fact shows the non-uniformity of the polystyrene surface in terms of cell culture. There is at present no simple test to monitor that surface. This paper presents a novel biologically based assay to determine the degree of heterogeneity of flasks supplied by various manufacturers. RESULTS: This paper shows that significant variation exists in polystyrene surface heterogeneity among several brands of tissue culture flasks, varying from 4 to 20% of the flask surface. There is also large variability within the production lot of a manufacturer. The assay method involves loading the cells with a cytoplasmic fluorescent marker that is released upon cell membrane rupture. Cell membrane rupture also causes the loss of marker proteins such as GAPDH used in Westernblots. This novel assay method can be used to monitor the batch consistency and the manufacturing process of flasks/dishes. It may also be used to test new biomaterials.