A TECHNIQUES FOR THE CULTURE OF BARRETTS ESOPHAGEAL CELLS

Establishment of cells in tissue culture from Barrett's columnar epith elium has been difficult. The aim of this study was to develop a succe ssful tissue culture method employing a serum-free medium for cultivat ion of Barrett's epithelial cells. Fragments of Barrett's mucosal tiss ue were explanted in a 3:1 mixture of Dulbecco's modification of Eagle 's medium and Ham's F12, to initiate the outgrowth of epithelial cells . Subsequently, a commercial serum-fret medium (formulated for the gro wth of keratinocytes) was used for the propagation of Barrett's oesoph agus cells without fibroblast growth. Cells established in culture ret ained their epithelial morphology, stained positive for cytokeratin, a nd contained Alcian blue (pH 2.5) and periodic acid-Schiff reagent-pos itive/diastase-resistant vacuoles, confirming their origin from Barret t's epithelium. Electron microscopy showed tonofilaments, microvilli a nd desmosomes. Coating the surface of culture vessels was not required and four cell strains could be passaged up to 20 times with no fibrob last growth, in the keratinocyte serum-free medium.