Yanbian Dairy Goat Mammary Wpithelial Cell Isolation, Culture And Identification

This test uses tissue culture method and the continuous passage law Yanbian dairy goat mammary epithelial cells in primary culture, purification and subculture, and by cell viability assay, growth curve drawing, karyotype analysis, oil red O staining, G418minimum lethal concentration determination, plasmid amplification and restriction enzyme digestion, cell transfection and other technical means to identify the cells, and fibroblasts Yanbian dairy goat comparison. The test results showed that the cell was a typical S-shaped growth curve, the number of chromosome60, cytoplasmic lipid droplets visible mammary epithelial cells by oil red staining. Indicates that the application of this test method for culturing cells for the Yanbian dairy goat normal mammary epithelial cells, and can secrete normal goat milk.1, Depending on the concentration of digestive enzymes can Yanbian dairy goat mammary epithelial cells and fibroblasts isolated and purified both get two pure cell lines.2, The application of two cryopreservation solution with10%DMSO15%glycerol cells were cryopreserved cells were thawed after cryopreservation solution on both adherent no difference in the rate of cells.3, The application of DMSO frozen dairy goat mammary epithelial cell protective agent, add frozen effect30%-50%FBS no significant difference in cells, the cells can be used as long-term preservation of frozen liquid.4, With added D-Hanks’formulated Yanbian0.15%trypsin digestion of dairy goat mammary epithelial cells recovered0.02%EDTA [48,73,79], after its passage grow better.5, Add hydrocortisone and epidermal growth factor to10%FBS in DMEM culture medium, more suitable for dairy goats Yanbian purified mammary epithelial cell growth.