动物造模 z-bio 2021-01-22 271

　　OBJECTIVE: To establish a method to isolate high-purity pulmonary microvascular endothelial cells, and to observe the wild-type mouse cells and receptors in knockout mice with high glycation end products stimulated by lipopolysaccharide.

　　Method: Collect the lungs of wild-type C57 mice and RAGE knockout mice aged 6 to 8 weeks, digest with type I collagenase, filter with nylon cells, and isolate the lungs of primary mice by the immunomagnetic bead method. Microvascular endothelial cells (PMVEC) were identified by morphology and immunofluorescence, and LPS (1 mg/L) was used to stimulate different time periods. The changes of cytoskeleton F-actin were observed with a laser confocal microscope.

　　Result: Under the microscope, it can be seen that the PMVEC that was cultured for the first time is spindle-shaped or polygonal, and it is highly purified by staining the cells against factor VIII-related antigen (VIIIF-Ag). After LPS stimulation, the PMVEC skeleton of wild-type mice will rearrange and form stress. The fiber PMVEC backbone of AGE knockout mice was not damaged.

　　 Conclusion: High-purity mouse lung microvascular endothelial cells can be obtained through immunomagnetic beads. RAGE is involved in the destruction of mouse lung microvascular endothelial cytoskeleton mediated by LPS.