高级微生物课 论文讲解概要课件.ppt

* * * 修磊 微生物 Abstract Introduction Materials and Methods Results Discussion Table Supplemental Ethics Statement Primary Bovine Mammary Epithelial Cell Culture Photomicrographs of the process obtained bovine epithelial cells and morphology of epithelial cells. (A-F). In panel A, single cells and small epithelioid islands of cells were apparent, spread upon the substrate after 24 h later, single cells with the granular appearance of macrophages were present. which noted by red color (100×); In panel B, a large epithelioid islands generated from a single cell after 3 days , Occasional macrophages were present,which made up 5% of all attached cells (100×); . In panel C, many large epithelioid islands generated were present after 9 days , and phagocytes were disappeared (100×); In panel D, areas of large cells arose within islands after two weeks in culture (100×); In panel E, the emulsion droplets secreted to the outside of the cells(100×); In panel F, the purified epithelial cells obtained over three passages(100×). Immunofluorescence analysis of bovine mammary cells (G-H). In panel G, first antibody (anti-cytokeratin 8) replaced by PBS as negative control, which pictured by fluorescence microscope. The cell nucleus were blue (DAPI) (100×); In panel H, fluorescent image of cells incubated with anti-cytokeratin 8 monoclonal antibody, pictured by fluorescence (100×). Invasion Assay PBMECs were washed three times. Thereafter, 1 ml of a suspension of S. aureus in DMEM was added per well. Cell monolayers infected with bacteria were incubated at37℃ in 5% CO2for 4 h. After incubation, monolayers were washed three times with 1 ml/well of phosphate buffered saline (PBS, pH 7.4), supernatants were replaced with tissue-growth medium containing gentamicin (100 mg/ml) and incubated for2 h at 37℃. After cell-culture medium containing antibiotics was re