已经开发了几种人工尿液(AU)配方来模拟正常尿液。它们中的大多数是无蛋白质的,特别是当分泌的蛋白质(分泌组)被分析。然而,正常的尿液实际上含有少量的蛋白质。我们假设生理水平的尿蛋白在保留肾细胞生物学和功能中起作用。这项研究评估了将0-10%胎牛血清(FBS)补充到完善的AU-Siriraj方案对MDCK肾小管细胞的影响。与完全培养基(对照)相比,天然尿液和不含/不含FBS的AU-Siriraj都减少了变形时间(TD)。在没有/没有FBS的天然尿液和AU-Siriraj中,AU-Siriraj+2.5%FBS中的细胞具有最长的TD。补充FBS以剂量依赖性方式增加细胞死亡(但仍<10%)。天然尿液中极化细胞的跨上皮电阻(TER)与对照相当,而AU-Siriraj+2.5%FBS中的细胞具有最高的TER。这些数据表明,向AU-Siriraj中补充2.5%FBS可以延长变形时间并增强肾小管细胞的极化。因此,AU-Siriraj+2.5%FBS强烈建议用于细胞生物学和功能的体外研究(当不对分泌组进行分析时)。
Several artificial urine (AU) formulas have been developed to mimic the normal urine. Most of them are protein-free, particularly when secreted proteins (secretome) is to be analyzed. However, the normal urine actually contains a tiny amount of proteins. We hypothesized that urinary proteins at physiologic level play a role in preservation of renal cell biology and function. This
study evaluated the effects from supplementation of 0-10% fetal bovine serum (FBS) into the well-established AU-Siriraj protocol on MDCK renal tubular cells. Time to deformation (TD) was reduced by both native urine and AU-Siriraj without/with FBS compared with complete culture medium (control). Among the native urine and AU-Siriraj without/with FBS, the cells in AU-Siriraj+2.5% FBS had the longest TD. Supplementation of FBS increased cell death in a dose-dependent manner (but still <10%). Transepithelial electrical resistance (TER) of the polarized cells in the native urine was comparable to the control, whereas that of the cells in AU-Siriraj+2.5% FBS had the highest TER. These data indicate that supplementation of 2.5% FBS into AU-Siriraj can prolong time to deformation and enhance polarization of renal tubular cells. Therefore, AU-Siriraj+2.5% FBS is highly recommended for in vitro
study of cell biology and function (when secretome is not subjected to analysis).