Cultures of exfoliated epithelial cells from different locations of the human urinary tract and the renal tubular system
Cultures of exfoliated epithelial cells from different locations of the human urinary tract and the renal tubular system
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DOI：
10.1007/s002040000173
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发表时间：
2000-12-01
影响因子：
6.1
通讯作者：
Föllmann, W
中科院分区：
文献类型：
--
作者：
Dörrenhaus, A;Müller, JIF;Föllmann, W
从健康成人、泌尿外科患者和内科患者的尿沉渣中分离并培养脱落的人尿路上皮细胞和肾小管细胞。细胞在接种沉淀物后1周内开始增殖。增殖细胞形成不同形态的集落，称为1型或2型细胞集落。1型细胞集落显示出不规则的轮廓和集落内的梭形细胞。1型细胞传代培养最多6代是可能的。2型细胞集落显示边缘光滑的轮廓，不可能进行传代培养。细胞角蛋白7免疫组化染色呈阳性，表明1型细胞的上皮特征。与碳酸酐酶阳性染色的Madin达比犬肾细胞（MDCK）（用作肾小管细胞的阳性对照）相反，1型细胞在磷酸钴法染色时为碳酸酐酶阴性。这表明1型细胞不是肾小管来源的。2型细胞碳酸酐酶染色阳性，表明2型细胞为肾小管细胞。2型细胞集落可分为两个亚组，具有不同的细胞形式。鹅卵石样细胞的集落比梭形细胞的2型细胞集落更常见，这是本研究首次描述。具有鹅卵石样细胞的集落形成圆顶（半囊），而梭形样2型细胞集落没有。比较了健康成人、接受呋塞米治疗的老年多病患者和接受磺胺甲恶唑/甲氧苄啶和/或经皮肾造瘘导管治疗的泌尿系结石患者的尿沉渣培养物。在52%的健康成人的所有培养沉积物中，在30%的多病患者中，在75-80%的泌尿科患者中，细胞增殖成集落。1型与2型细胞集落的比例分别为3.3：1（健康成人）、1.4：1（泌尿系结石患者）和1.8：1（泌尿系结石患者，使用经皮肾造口术导管直接从肾盂收集尿液）。这三组尿沉淀物的成功培养分别显示平均3或4个菌落、14个菌落和21个菌落。仅在泌尿科患者组中观察到与性别有关的菌落数差异。结果表明，1型细胞是尿路上皮细胞，由于它们在尿路内的起源位置，没有显示出形态学差异，而2型细胞可能是肾小管细胞。这些发现提供了新的方面，在培养人尿路上皮或肾上皮细胞的方法的基础上，无创收集标本，只需要最小的培养工作。通过该方法获得的培养物可用于毒理学和临床研究中的体外研究。
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.