INFLUENCES OF EXTRACELLULAR-MATRIX COMPONENTS ON THE GROWTH AND DIFFERENTIATION OF RUMINAL EPITHELIAL-CELLS IN PRIMARY CULTURE

Primary cultures of ovine ruminal epithelial cells were made to study the influence of collagen types I and IV and of medium supplementation with various hormones and Na-n-butyrate on cell morphology and growth characteristics. Both collagen type I and type IV led to increased cell proliferation with the stimulatory effect being more pronounced in collagen IV. In cultures grown on collagen I, both non-stratified and stratified colonies were found, whereas cultures grown on collagen IV showed predominantly stratified growth. Cells in both stratified and non-stratified colonies were positive for cytokeratin antibody. In non-stratified colonies, positive staining with fibronectin antibodies (FN-15) was found in a network over and around the cells. It is suggested that the non-stratified ruminal epithelial cells are in some respects similar to a 'non-differentiating keratinocyte' strain, derived from newborn foreskin epidermis. Cells in stratified colonies bound Ulex europaeus (UEAI) lectin which has been shown to be specific for differentiated epithelial cells in ruminal mucosa. Supplementation of culture medium with glucagon and insulin increased the total cell-overgrown area of collagen I cultures, whereas this effect was absent in cultures grown on collagen IV. In both cultures grown on collagen I or IV, hydrocortisone led to an increase in total cell-overgrown area, whereas Na-n-butyrate inhibited proliferation.