Improvements in Culturing Exfoliated Urothelial CellsIn Vitrofrom Human Urine

Abstract

Human bladder cancer is a common malignant tumor that may be produced by factors such as lifestyle, environment and occupation. The aim of this study was to evaluate parameters related to the viability of exfoliated urothelial cells. Exfoliated urothelial cells were obtained from 83 urine samples of 22 healthy participants (20-53 yr). From 67 of these samples, cells were transferred to collagen-coated 24-well plates. Parameters including sample volume, pH, osmolality and participant age and gender were examined on cell viability. In successive cultures, the numbers of cell colonies and cells per cell colony were determined. The number of viable cells in the urinary sediments of males varied from 0 to 6.5 x 10(3) cells per sample (mean 1 x 10(3)). Higher cell numbers in urine samples from females (6 x 10(3)) were due to considerable amounts of exfoliated vaginal cells. Cell numbers in males were positively related to volume, osmolality, and pH of the samples, as well as to the retention time of urine in the bladder. Cell proliferation was achieved in 25 out of 67 samples and was positively related to sample osmolality and pH. Participant age and content of urinary oxalates exerted negative effects on cell proliferation in vitro. The mean number of cell colonies per sample was 1.7. The mean cell number per colony was 11.7 x 10(3). It appears that high variability in individual excretion of urothelial cells able to proliferate is a limiting factor for routine use of these cells for in vitro toxicology.

Urine is a novel source of autologous mesenchymal stem cells for patients with epidermolysis bullosa

“…As expected, the number of colonies was significantly higher when the plates were not coated ( p = 0.026) (Fig. 2 b), since adherence to plastic surfaces is typical for mesenchymal stem cells [ 14 ].…”

“…Urine pH was measured and samples from healthy donors were also evaluated for the presence of glucose, leucocytes, nitrites, proteins and blood using Combur Test HC (Cobas) [ 14 ].…”

Adenoviral infectivity of exfoliated viable cells in urine: Implications for the detection of bladder cancer

“…In agreement with a recent study we found that cell attachment and propagation occurred within 10 days after culture initiation [17]. About half of all samples (16/31) showed evidence of in vitro growth.…”

Advances in understanding the cell types and approaches used for generating induced pluripotent stem cells

“…Therefore, urine samples may be considered a preferred source for iPSC derivation. However, other reports indicate that the number of excreted cells range from 0 to 6500 showing a high degree of individual variation, and cell proliferation is achieved in only approximately 1/3 of the samples [[ 55 ]]. This raises the questions whether HUCs can be universally applied for generating iPSCs.…”

“…The reprogramming efficiency of PBSs is extremely low (0.0008 to 0.001%), and the mature T cells derived iPSCs harbor germ line IgH and TCR alleles; moreover, in some cases of infection and blood diseases, giving blood is not exempt of concerns. HUCs seem to be more promising than PBSs, they are non-invasive and can be reprogrammed with a high efficiency even with an integration-free reprogramming method (0.2%), However, other reports indicate a high degree of variability of excreted cell number in urine among individuals, and the proliferation was achieved in only approximately 1/3 of samples [[ 55 ]]. Moreover, culturing HUCs usually requires long periods of time (at least two weeks) [[ 53 ]].…”