Cultures of exfoliated epithelial cells from different locations of the human urinary tract and the renal tubular system

Abstract

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.

Glucose Transporters in Human Renal Proximal Tubular Cells Isolated From the Urine of Patients With Non–Insulin-Dependent Diabetes

“…1D). Two types of colonies arose: type 1 colonies, with irregular contours, and type 2 colonies, with smooth edges, were both observed as described by Dorrenhaus et al (24). The cells were characterized by Western blotting/ Taqman RT-PCR and confocal microscopy ( Table 1) and proved to be a heterogeneous population as also found by Inoue et al (23), who grew mixed primary cultures of distal and proximal cells from voided urine on glass coverslips.…”

“…In contrast, culture of specific renal tubular cells from human urine could offer a noninvasive and novel tool for renal research in humans. However, only 2,000 -7,000 renal cells of various types are normally exfoliated each day (21), and this has meant that only few groups have successfully cultured mixed renal tubular cells from urine (22)(23)(24). To our knowledge, no one has reported the successful isolation and subculture of enriched populations of proximal tubular cells from the urine of healthy and type 2 diabetic subjects to date.…”

Decursin Prevents Cisplatin-Induced Apoptosis via the Enhancement of Antioxidant Enzymes in Human Renal Epithelial Cells

“…Cell Culture Primary HRCs isolated from the urine of a healthy, non-stone forming male using the method of Dorrenhaus 12) were gratefully obtained from Dr. John C. Lieske. 13) Cells were maintained in Dulbecco's modified Eagle's medium (DMEM) (Welgene, Deagu, South Korea) supplemented with fetal bovine serum (FBS) and 100 units/ml penicillin and 100 mg/ml streptomycin at 37°C in a humidified atmosphere containing 5% CO 2 .…”

Generation of Induced Pluripotent Stem Cells from Urine

“…These colonies corresponded to two main morphologies: type 1 or type 2 ( Figure 1B), in agreement with previous reports on urine cell isolation. 18 Type 1 cells were more rounded and grew closely attached to neighbor cells, suggesting an epithelial phenotype. Type 2 cells were more elongated and grew more dispersed.…”