ABMES

Non-adherent cell culture surface has been widely used for producing cell spheroids and cell aggregates. The purpose of this study was to formulate a new method for non-adherent cell culture without coating or surface-modification that has been needed. We found that high-molecular-weight dextran sulfate (DS) and bovine serum albumin (BSA) synergistically prevented cell adhesion in media supplemented with no or low serum. This method worked on tissue culture-treated polystyrene surfaces as well as on commercially available low-attachment- and untreated polystyrene surfaces. Further investigation revealed that BSA may mediate the adsorption of DS to the surface. In addition, as the adsorption of fluorescently labeled fibronectin was inhibited by BSA alone, it appears that protein adsorption and cell adhesion do not always correlate. Finally, we demonstrated the successful formation of HepG2 spheroids and cardiomyocyte aggregates using this method. In conclusion, cell adhesion can be effectively suppressed by simply adding DS and BSA to the culture medium without coating or surface modification, and it may be useful for generating cell spheroids and aggregates.