本研究的目的是比较四种不同的尿路上皮细胞分离和培养方法,并将其与文献中引用的方法进行比较。检查了四种不同的技术以从大鼠膀胱中分离尿路上皮。使用锥虫蓝测定法计算分离有效性。使用免疫组织化学(IHC)确认分离的细胞表型并与天然膀胱组织进行比较。免疫细胞化学(ICC)和免疫荧光(IF)分析。膀胱倒置和胶原酶P消化的方法导致最高数量的分离细胞。这些细胞显示细胞角蛋白7、8、18、α6-整联蛋白和p63的阳性表达。我们的结果和文献综述表明,膀胱尿路上皮分离的最佳方法是从其他膀胱部位解剖上皮层,并在胶原酶溶液中消化机械制备的组织。
The aim of this study is to present the comparison of four different methods for urothelial cell isolation and culture and compare them to methods cited in the literature. Four different techniques were examined for urothelium isolation from rat bladders. Isolation effectiveness was calculated using trypan blue assay. Confirmation of isolated cell phenotype and comparison with native bladder tissue was confirmed using immunohistochemical (IHC), immunocytochemical (ICC) and immunofluorescence (IF) analysis. The method with bladder inversion and collagenase P digestion resulted in the highest number of isolated cells. These cells showed positive expression of cytokeratin 7, 8, 18, α6-integrin and p63. Our results and the literature
review showed that the best method for urothelium bladder isolation is dissection of the epithelium layer from other bladder parts and digestion of mechanically prepared tissue in a collagenase solution.