Dörrenhaus A, Müller J I, Golka K, Jedrusik P, Schulze H, Föllmann W
Institut für Arbeitsphysiologie an der Universität Dortmund, Germany.
Arch Toxicol. 2000 Dec;74(10):618-26. doi: 10.1007/s002040000173.
Exfoliated human urinary tract epithelial cells and renal tubular cells from urinary sediments of healthy adults, of urological patients and of internal patients were isolated and cultured. Cells started proliferating within 1 week after seeding a sediment. Proliferating cells formed colonies of different morphologies, designated as type-1 or type-2 cell colonies. Type-1 cell colonies showed irregular contours and spindle-like cells within the colonies. Subcultivation of type-1 cells for up to six passages was possible. Type-2 cell colonies showed smooth-edged contours and subcultivation was not possible. The epithelial character of type-1 cells was demonstrated by positive immunohistochemical staining for cytokeratin-7. In contrast to carbonic anhydrase-positive stained Madin Darby canine kidney cells (MDCK), which were used as positive controls for renal tubular cells, type-1 cells were carbonic anhydrase-negative on staining with the cobalt phosphate method. This indicates that type-1 cells were not of renal tubular origin. Type-2 cells were positively stained for carbonic anhydrase, indicating that type-2 cells were renal tubular cells. Type-2 cell colonies could be assigned to two subgroups with different cell forms. Colonies of cobblestone-like cells more often occurred than type-2 cell colonies with spindle-like cells, which are described in this study for the first time. Colonies with cobblestone-like cells formed domes (hemicysts), whereas spindle-like type-2 cell colonies did not. Cultures of urinary sediments from healthy adults, elderly multimorbid patients treated with furosemide, and urological patients with urolithiasis treated with sulfamethoxazole/trimethoprim and/or with a percutaneous nephrostomy catheter were compared. In 52% of all cultured sediments from healthy adults, in 30% of those from multimorbid patients, and in 75-80% of those from urological patients cells proliferated to colonies. The ratios of type-1 to type-2 cell colonies were 3.3:1 (healthy adults), 1.4:1 (urological patients with urolithiasis), and 1.8:1 (urological patients with urolithiasis, urine was directly collected from the renal pelvis with a percutaneous nephrostomy catheter). Successful cultures of the urinary sediments from these three groups revealed means of 3 or 4 colonies, 14 colonies, and 21 colonies, respectively. Differences in the number of colonies in relation to sex were observed only for the group of urological patients. It was shown that type-1 cells were urothelial cells, which did not show morphological differences due to their locations of origin within the urinary tract, whereas type-2 cells were probably renal tubular cells. These findings offer new aspects in the culturing of human urothelial or kidney epithelial cells with a method based on noninvasive collecting of specimens and requiring only minimal culture effort. The cultures obtained by this method can be used for in vitro studies in toxicological and clinical research.
从健康成年人、泌尿系统疾病患者和内科患者的尿沉渣中分离并培养了脱落的人尿道上皮细胞和肾小管细胞。接种尿沉渣后1周内细胞开始增殖。增殖的细胞形成了不同形态的集落，分为1型或2型细胞集落。1型细胞集落轮廓不规则，集落内有梭形细胞。1型细胞可传代培养多达6代。2型细胞集落轮廓边缘光滑，无法传代培养。1型细胞的上皮特征通过细胞角蛋白-7免疫组织化学染色阳性得以证明。与用作肾小管细胞阳性对照的碳酸酐酶染色阳性的Madin Darby犬肾细胞（MDCK）相比，1型细胞用磷酸钴法染色时碳酸酐酶呈阴性。这表明1型细胞不是肾小管来源。2型细胞碳酸酐酶染色呈阳性，表明2型细胞是肾小管细胞。2型细胞集落可分为两个具有不同细胞形态的亚组。鹅卵石样细胞的集落比首次在本研究中描述的梭形2型细胞集落更常见。鹅卵石样细胞的集落形成穹顶（半囊肿），而梭形2型细胞集落则不形成。比较了健康成年人、接受速尿治疗的老年多病患者以及接受磺胺甲恶唑/甲氧苄啶和/或经皮肾造瘘导管治疗的尿路结石泌尿系统疾病患者的尿沉渣培养物。在所有来自健康成年人的培养尿沉渣中，52%的尿沉渣中有细胞增殖形成集落；来自多病患者的尿沉渣中，这一比例为30%；来自泌尿系统疾病患者的尿沉渣中，这一比例为75 - 80%。1型与2型细胞集落的比例分别为3.3:1（健康成年人）、1.4:1（尿路结石泌尿系统疾病患者）和1.8:1（尿路结石泌尿系统疾病患者，经皮肾造瘘导管直接从肾盂收集尿液）。这三组尿沉渣的成功培养分别显示平均有3或4个集落、14个集落和21个集落。仅在泌尿系统疾病患者组中观察到集落数量与性别有关的差异。结果表明，1型细胞是尿道上皮细胞，因其在尿道内的起源位置不同而未表现出形态差异，而2型细胞可能是肾小管细胞。这些发现为基于非侵入性标本采集且仅需极少培养工作的方法培养人尿道上皮或肾上皮细胞提供了新的视角。通过这种方法获得的培养物可用于毒理学和临床研究的体外研究。
