How to isolate urothelial cells? Comparison of four different methods and literature review

Kloskowski, Tomasz; Uzarska, Małgorzata; Gurtowska, Natalia; Olkowska, Joanna; Joachimiak, R.; Bajek, Anna; Gagat, Maciej; Grzanka, Alina; Bodnar, Magdalena; Marszałek, Andrzej; Drewa, Tomasz

doi:10.1007/s13577-013-0070-y

Cited by 24 publications

(31 citation statements)

References 32 publications

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“…In Vitro Experiments NLRP3 +/+ , nondiab mice (i.e., control littermates) were used at 7 to 8 weeks of age. Urothelial cells were isolated (26) and plated (black-walled 96-well plates) at 50,000 cells/well in 90 mL complete media (F-12K media, 10% lowendotoxin dialyzed FBS, 10 mmol/L nonessential amino acids [all from HyClone Laboratories, Logan, UT], 1.0 mg/mL hydrocortisone [Sigma-Aldrich, St. Louis, MO], 10 mg/mL insulin, 5 mg/mL transferrin, and 6.7 ng/mL selenium [Gibco, Gaithersburg, MD]). Following a 24-h incubation (37°C, 95% air/5% CO 2 ), DAMPs (10 mL) were added and incubated as indicated.…”

Section: Animalsmentioning

confidence: 99%

NLRP3 Promotes Diabetic Bladder Dysfunction and Changes in Symptom-Specific Bladder Innervation

et al. 2018

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The NLRP3 inflammasome senses diabetic metabolites and initiates inflammation implicated in diabetic complications and neurodegeneration. No studies have investigated NLRP3 in diabetic bladder dysfunction (DBD), despite a high clinical prevalence. In vitro, we found that numerous diabetic metabolites activate NLRP3 in primary urothelial cells. In vivo, we demonstrate NLRP3 is activated in urothelia from a genetic type 1 diabetic mouse (Akita) by week 15. We then bred an NLRP3 2/2 genotype into these mice and found this blocked bladder inflammation and cystometric markers of DBD. Analysis of bladder innervation established an NLRP3-dependent decrease in overall nerve density and Ad-fibers in the bladder wall along with an increase in C-fiber populations in the urothelia, which potentially explains the decreased sense of bladder fullness reported by patients and overactivity detected early in DBD. Together, the results demonstrate the role of NLRP3 in the genesis of DBD and suggest specific NLRP3-mediated neuronal changes can produce specific DBD symptoms.

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Section: Animalsmentioning

confidence: 99%

NLRP3 Promotes Diabetic Bladder Dysfunction and Changes in Symptom-Specific Bladder Innervation

et al. 2018

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“…There are a number of different ways to isolate and culture human urothelial cells . However, obtaining urological tissue for primary urothelial cell cultures is not feasible, since no healthy subject would provide informed consent for the required hospitalisation and handling for the donation of bladder tissue .…”

Section: Discussionmentioning

confidence: 99%

“…28 There are a number of different ways to isolate and culture human urothelial cells. 29 However, obtaining urological tissue for primary urothelial cell cultures is not feasible, since no healthy subject would provide informed consent for the required hospitalisation and handling for the donation of bladder tissue. 30 It is possible to culture urothelial cells from bladder washing or donated urine, but such cell cultures only retain their stability for a few passages and they quickly stop proliferating.…”

Section: Discussionmentioning

confidence: 99%

Artificial urine and FBS supplemented media in cytocompatibility assays for PLGA‐PEG‐based intravesical devices using the urothelium cell line UROtsa

Arndt

Leistner²,

Neuß

et al. 2017

J Biomed Mater Res

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European and German directives for approval of new medical devices require tests for cytotoxicity in relevant media, since urine can influence cytotoxicity of biodegradable devices. The aim of this study was to determine the long-term cytotoxicity of PLGA-b-mPEG (PLGA-PEG) polymer carriers and artificial urine (AU) to human UROtsa cells. Benign urothelial UROtsa cells were incubated in fetal bovine serum-containing RPMI 1640 medium supplemented with a range of concentrations of AU for 24 h and 7 days. Cell viability was determined by the XTT assay and by live/dead staining. The cytotoxicity of medium containing degradation products from PLGA-PEG carriers was also tested on the UROtsa cells in AU-containing and control medium. PLGA-PEG carriers exhibited no cytotoxicity to UROtsa cells after 24 h of incubation. However, after 7 days, cytotoxicity was observed, but this was largely attributable to the effects of 30% AU on the cells. Compared to phosphate buffer saline (PBS) and normalized to RPMI 1640 medium, significant cytotoxicity was observed by 24 h in medium containing 50% AU and by 7 days in medium containing 30% AU. Live/Dead staining confirmed proliferation results and no pH-changes could be observed. Here we demonstrate for the first time the impact of AU on standard cytotoxicity tests related to biomaterials for urinary-tract applications. Our study showed cytotoxic effects of high concentrations of 50% AU by 24 h and by physiological concentrations of AU (i.e., 30%) by 7 days. We have also demonstrated that PLGA-PEG has no cytotoxic effects in the appropriate AU-containing test environment. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2140-2147, 2018.

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“…Some of these drugs showed the activity through inhibition of mammalian DNA topoisomerases in addition to other mechanisms (Figure ) . Ciprofloxacin, as an example, revealed a considerable activity and induced apoptosis in different cancer cell lines . It showed the advantage of inducing apoptosis and cell death in malignant cells but not in normal ones and giving a suggestion to be used as an adjuvant therapy for the treatment of lung cancer .…”

Section: Antibacterial Fluoroquinolone Drugs With Anticancer Activitymentioning

confidence: 99%

Towards anticancer fluoroquinolones: A review article

Abdel-AAl

Abdel‐Aziz

Shaykoon

et al. 2019

Archiv der Pharmazie

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Different studies about the anticancer potential of several medically used antibacterial fluoroquinolones have been established. Fluoroquinolone derivatives, like some anticancer drugs, such as doxorubicin, can achieve antitumor activity via poisoning of type II human DNA topoisomerases. Interestingly, structural features required for the anticancer activity of quinolones have been determined. Most of the chemical modifications required to convert antibacterially acting fluoroquinolones into their anticancer analogs were at position 7 and the carboxylic group at position 3. This review highlights the antitumor potential of fluoroquinolones in general and summarizes the chemical modifications carried out on fluoroquinolones to become anticancer agents. Moreover, the review gives a quick recap on metal ion chelates with fluoroquinolones and their substantial role in topoisomerase poisoning and antitumor potential improvement. Hence, it should be highly interesting for researchers attempting to design and synthesize novel anticancer fluoroquinolone candidates. K E Y W O R D S anticancer, DNA topoisomerase, metal ion complexes, quinolones Arch Pharm Chem Life Sci. 2019;352:1800376.wileyonlinelibrary.com/journal/ardp

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How to isolate urothelial cells? Comparison of four different methods and literature review

Cited by 24 publications

References 32 publications

NLRP3 Promotes Diabetic Bladder Dysfunction and Changes in Symptom-Specific Bladder Innervation

NLRP3 Promotes Diabetic Bladder Dysfunction and Changes in Symptom-Specific Bladder Innervation

Artificial urine and FBS supplemented media in cytocompatibility assays for PLGA‐PEG‐based intravesical devices using the urothelium cell line UROtsa

Towards anticancer fluoroquinolones: A review article

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