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.

非贴壁细胞培养表面已广泛用于生产细胞球体和细胞聚集体。本研究的目的是制定一种新的非贴壁细胞培养方法，无需涂层或表面改性。我们发现高分子量的硫酸葡聚糖 (DS) 和牛血清白蛋白 (BSA) 在补充无血清或低血清的培养基中协同防止细胞粘附。这种方法适用于组织培养处理的聚苯乙烯表面以及市售的低附着和未处理的聚苯乙烯表面。进一步的研究表明，BSA 可能介导 DS 对表面的吸附。此外，由于荧光标记的纤连蛋白的吸附被单独的 BSA 抑制，因此蛋白质吸附和细胞粘附似乎并不总是相关。最后，我们证明了使用这种方法成功形成了 HepG2 球体和心肌细胞聚集体。总之，只需在培养基中加入 DS 和 BSA 即可有效抑制细胞粘附，无需涂层或表面修饰，可用于生成细胞球体和聚集体。