METHYLCELLULOSE PROTECTS THE ABILITY OF ANCHORAGE-DEPENDENT CELLS TO ADHERE FOLLOWING ISOLATION AND HOLDING IN SUSPENSION

Preparation of suspension of anchorage-dependent cells growing in tiss ue cultures requires removal of the cells from their substrata by mean s of trypsin and/or EDTA. The purpose of the experiments was to invest igate the effect of methylcellulose on the adhesive ability of cells r emoved from substrata by EDTA and trypsin. Human umbilical cord vein e ndothelial cells (HUVEC), after detachment from substrata, adhere well to fibronectin at 37 degrees C but not at 0 degrees C. During a 60-mi n incubation at 37 degrees C, these cells lose about 70% of their abil ity to adhere to fibronectin. The ability of cells to adhere was resto red in the presence of 0.2% methylcellulose. Methylcellulose also prev ented human skin fibroblasts, human melanoma cells and mouse lung fibr oblasts from losing adhesive properties. By contrast, it did not affec t the adhesive ability of B16F10 melanoma cells incubated at 37 degree s C. In conclusion, our study suggests that methylcellulose can be a u seful reagent for preservation of cell function in suspension; it may also simplify some experimental procedures including radioiodination o f cell surface components and cross-linking of radiolabeled ligands to the cell surface.