小鼠子宫巨噬细胞的分离纯化与鉴定

目的 建立一套简便易行、高效可靠的小鼠子宫巨噬细胞的分离纯化及鉴定方法. 方法 5只雌鼠处死并获取子宫,利用酶消化法和机械法相结合制备小鼠子宫组织单细胞悬液,密度梯度离心法收集子宫巨噬细胞,贴壁培养获得纯化的巨噬细胞.采用锥虫蓝拒染法鉴定巨噬细胞活率,利用鸡红细胞吞噬试验检测巨噬细胞吞噬活性,并通过酸性磷酸酶染色、非特异性酯酶染色和CD14免疫细胞化学染色鉴定巨噬细胞纯度. 结果 采用该法每只小鼠可获得子宫巨噬细胞3×105～5×105个,且活率达到(85.35±0.43)%,酶化学和免疫细胞化学鉴定阳性率分别达到了(86.12±1.19)%、(85.83±0.57)%和(84.02±1.64)%. 结论 成功建立了小鼠子宫巨噬细胞的分离纯化方法,并能获得较高的细胞存活率和纯度.

Isolation,purification and identification of macrophages from mouse uterus

Objective To establish an efficient and reliable method of isolating，purifying and identifying macrophages from mouse uterus. Methods Five female mice were killed and their uteri were obtained. Mechanical dissection and digestive enzymes were used to prepare cell suspension of mouse uterus. Uterine macrophages were separated by density gradient centrifugation. And these cells were purified by anchoring cultivation. Then the cell viability was assessed by trypan blue exclusion staining. The phagocytosis of cell was evaluated by phagocytosis of chicken erythrocytes. The purity of cell was determined by acid phosphatase staining，α acetic acid naphthol esterase staining and surface specific antigen CD14 staining. Results By using this new method，the cell viability was（85.35±0.43）％. And the purity of uterine macrophages was （86.12±1.19）%,（