title: Goblet cell isolation from cat trachea: a comparison of methods.
abstract: Goblet cell hyperplasia and metaplasia may be important in the pathogenesis of many respiratory diseases. To study the intracellular mechanisms of mucin synthesis, a purified goblet cell preparation is necessary. We have compared different methods of cell dissociation using cat trachea, a source rich in goblet cells. The most successful method used EDTA to loosen the basement membrane and 1% pronase to dissociate the epithelial cells. Goblet cells recovered from a linear Percoll gradient showed preservation of their ultrastructural detail, trypan blue exclusion, oxygen consumption, and synthesis of a high molecular weight compound resistant to hyaluronidase degradation, consistent with mucous glycoprotein. This method allows the isolation of adequate numbers of purified goblet cells for further study of goblet cell synthetic processes.
mesh_terms: D000818:Animals; D002415:Cats; D002469:Cell Separation / Q000379:methods*; D004492:Edetic Acid; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D009077:Mucins / Q000096:biosynthesis; D011402:Pronase; D014132:Trachea / Q000166:cytology* / Q000378:metabolism
chemical_list: D009077:Mucins; D004492:Edetic Acid; D011402:Pronase