PRIMARY CULTURE OF BOVINE GALL-BLADDER EPITHELIAL-CELLS

Gall bladder epithelial cells serve numerous biological functions and abnormalities in their function are important in the pathogenesis of s everal gall bladder diseases. Direct studies on cell function are rare due to lack of reliable methods to culture this epithelium. This stud y reports a reliable and reproducible method of harvesting and culturi ng gall bladder epithelial cells. Normal bovine gall bladder epitheliu m, obtained within 20 minutes of slaughter, was rinsed with modified H anks's balanced salt solution, the mucosa separated and incubated in t rypsin - EDTA solution at 37-degrees-C. The cells were isolated and re suspended in Dulbeco's modification of Eagles' medium containing 10% f etal calf serum and, after filtration and centrifugation, were plated under aspetic conditions. The growth rate was established by flow cyto metry and the morphological characteristics of the growing cells by el ectron microscopy. Gall bladder epithelial cells grew successfully and visible clusters of cells were present by day two, confluency being r eached at 8 to 10 days in collagen coated plates and 12 to 14 days in uncoated plates. Electron microscopy showed typical gall bladder epith elia with microvilli, tight junctions, and mucus droplets. This method proved reliable and reproducible tor the culture of gall bladder epit helial cells and should allow direct studies of the biological propert ies of these cells in human tissue.