Urinary Tract Cancer

In this video, we demonstrate the vibratome sectioning of a whole mouse kidney to obtain thin slices. These sliced tissue sections can be stored at low temperatures until further downstream analysis.

In this video, we demonstrate SPME-probe-based extraction of metabolites from a pre-harvested donor kidney. The extracted metabolites can be used to perform metabolomic and lipidomic analysis in order to assess the organ quality for transplantation.

This video describes the technique of staining extracellular DNA in FFPE tissue sections. The stained extracellular DNA can be quantitated to determine the efficacy of any therapeutic treatment that leads to the death of cancer cells.

In this video, we describe the protocol to derive human kidney podocytes from induced pluripotent stem cells through cell differentiation under defined chemical conditions. The generated podocytes can be used for nephrotoxicity testing and disease modeling.

In this video, we demonstrate the isolation of detrusor smooth muscle cells from human urinary bladder specimens using a two-step sequential enzymatic treatment.

This video demonstrates the characterization of cationic currents from transient receptor potential melastatin type 4 (TRPM4) ion channels in detrusor smooth muscle cells using amphotericin-B mediated perforated patch-clamp technique. The technique helps evaluate the biophysical and pharmacological properties of ion channels.

This video describes the method to isolate intact glomeruli from mouse kidneys. The encapsulated and decapsulated glomeruli thus obtained can further be used for culturing parietal epithelial cell outgrowths from the murine kidney.

This video demonstrates the preparation of cancer spheroid blocks by flash freezing, a technique in which a tissue sample of interest is frozen to an ultracold temperature, restraining the spheroids inside the matrix.

This video describes a density-based ultracentrifugation technique for isolating exosomes from conditioned media harvested from human embryonic kidney cell cultures using a 25% (w/w) sucrose cushion prepared in deuterium oxide. The isolated exosomes can be used for downstream applications.

This video describes an electroporation-based method for encapsulating siRNA into exosomes. The siRNA-loaded exosomes can then be used for therapeutic applications.

In this video, we demonstrate the steps to isolate and purify renal tubular epithelial cells for a wide range of molecular biology and cell culture studies.

In this video, we describe the protocol for performing Periodic Acid Schiff (PAS) stain to assess the structural anomalies in the kidney glomerulus. The PAS staining method enables appropriate evaluation of the glomerulus pathophysiology.

This video demonstrates the methodology for performing Reverse Phase Protein Arrays (RPPA) to study protein expression patterns from cell lysate. In RPPA, the lysate containing a mixture of proteins is printed on nitrocellulose slides and proteins of interest are analyzed using fluorescently labeled antibodies.

In this video, we describe a method called expansion pathology, a variant of expansion microscopy, wherein the clinical tissue specimens are chemically expanded using a polymer network for microscopic examination of nanoscale structures.

This video describes the procedure to generate an experimental xenograft murine model, which can be used for studying urothelial cell carcinoma progression.

In this video, we describe the orthotopic transplantation of cultured bladder tumor organoids into the murine bladder to study the role of the organ environment on developing tumors.

This video demonstrates the procedure to generate a unilateral ureteral obstruction mouse model. The procedure involves cutting one of the ureters of the mouse to obstruct urine flow.

In this video, we describe the intravesical procedure of delivering cancer cells into the bladder of a mouse model. This procedure helps generate an orthotopic bladder tumor model that can be used to develop therapeutic strategies.

In this video, we describe a catheter-based procedure to empty and rinse the urinary bladder of a mouse model. The prepped mouse can then be used for further experimental procedures.

This video demonstrates the delivery of DNA plasmid into the urothelium of the mouse bladder using a combination of urethral catheterization and electroporation techniques. The described procedure allows the generation of mouse bladder cancer models.

In this video, we present the protocol for hydrodynamic injection of plasmids into the renal pelvis of the mouse kidney for nucleic acid transfection. This technique allows organ-specific delivery of plasmids by utilizing the principle of high hydrostatic pressure, which forcefully drives the plasmids into the specific site without causing tissue injury.

In this video, we develop an orthotopic mouse model of renal cell carcinoma. This model results in reproducible primary tumors in the injected kidney and metastatic tumors in the lung, allowing the evaluation of disease progression and treatment.

This video demonstrates the intravenous microinjection of nephrotoxins into the tail vein of a zebrafish embryo to study their chemical effects on renal function.

In this video, we demonstrate intra-renal drug administration in rats via the renal artery. This method is beneficial in improving drug targeting, drug potency, and minimizing systemic toxicity.

This video showcases fluid aspiration from mouse glomeruli using the two-photon mediated micropuncture technique. The technique provides access to the living mouse’s glomeruli and thus helps gain insights into renal physiology.

In this video, we describe the procedure for kidney extraction from the retroperitoneal space of the mouse. The extracted kidney can further be microdissected to obtain an intact kidney without a renal capsule and the surrounding fat layer.

This video describes the preparation of a decellularized rat kidney. The obtained acellular vascular scaffold can serve as a promising platform for tissue engineering of organ grafts.

This video demonstrates the dissection and preparation of a flat mount of the adult zebrafish kidney. The prepared flat mount can be visualized under the microscope for specimen analysis.

In this video, we demonstrate the depigmentation of adult zebrafish kidney and its subsequent preparation for staining procedures. Post-staining, the adult zebrafish kidney can be used for studying kidney regeneration, phenotypic characterization of renal disease models, and nephron functionality.

This video describes a method for studying organ development using an improved xenotransplantation setup on chorioallantoic membrane (CAM) from avian embryos. This method allows for the development of fully vascularized cultured embryonic organs and organoids.