Effects Of Goat Mammary Epithelial Cells On Peripheral Blood Polymorphonuclear Neutrophisls

Separated goat mammary epithelial cell with collagenase I and trypsin, gained in vitro culture. Tested the effect of goat mammary epithelial cell and its conditioned medium on PMN activity by MTT colorimetric assay, in order to illustrate the characteristic of mammary immune. The results as follow:1. Digesting goat mammary epithelial tissue with 2.0 g/L collagenase I for 4.5 h, 0.5 g/L trypsin for half an hour, gained more single cell than use collagenase only. Purifying the cell with 0.5 g/L trypsin&0.2 g/L EDTA for 3~4 times, identified the cells by immunohisto- chemistry, the purity rate of epithelia cell is more than 95%.2. Culturing mammary epithelial tissue with explant method, cells grow well on rat tail tendon collagen as well as without, this suggested that growth of mammary epithelial cell doesn't depend on rat tail tendon collagen.3. Groped three species segregation methods of PMN, Lymphocyte Separation Medium connected with percoll gradient centrifugation, erythrocyte physio-sedime-ntation acted by Dextran and dissolve erythroyte by tris-HCI. The result suggested that lymphocyte separation medium connected with percoll gradient centrifugation is the best way for the separation of PMN.4. Effect of goat mammary epithelial cell and its conditioned medium on PMN activity was detected by MTT colorimetric assay, the result showed that goat mammary epithelial cell and its conditioned medium have obvious inhibit effect on PMN. This suggested that mammary epithelial cell inhibits PMN fuction by some way.