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 tissue cultures requires removal of the cells from their substrata by means of trypsin and/or EDTA. The purpose of the experiments was to investigate the effect of methylcellulose on the adhesive ability of cells removed from substrata by EDTA and trypsin. Human umbilical cord vein endothelial cells (HUVEC), after detachment from substrata, adhere well to fibronectin at 37 degrees C but not at 0 degrees C. During a 60-min incubation at 37 degrees C, these cells lose about 70% of their ability to adhere to fibronectin. The ability of cells to adhere was restored in the presence of 0.2% methylcellulose. Methylcellulose also prevented human skin fibroblasts, human melanoma cells and mouse lung fibroblasts from losing adhesive properties. By contrast, it did not affect the adhesive ability of B16F10 melanoma cells incubated at 37 degrees C. In conclusion, our study suggests that methylcellulose can be a useful reagent for preservation of cell function in suspension; it may also simplify some experimental procedures including radioiodination of cell surface components and cross-linking of radiolabeled ligands to the cell surface.
Stewart
G J
GJ
Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA 19140, USA.
Wang
Y
Y
Niewiarowski
S
S
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects*; D002469:Cell Separation; D002478:Cells, Cultured; D004492:Edetic Acid; D004730:Endothelium, Vascular / Q000166:cytology; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D008168:Lung; D008545:Melanoma / Q000473:pathology; D008747:Methylcellulose / Q000494:pharmacology*; D051379:Mice; D012867:Skin; D014021:Tissue Preservation; D014357:Trypsin; D014407:Tumor Cells, Cultured; D014471:Umbilical Veins
D008747:Methylcellulose; D004492:Edetic Acid; D014357:Trypsin