Bladder Tumor Organoid Model: A Method for Orthotopic Transplantation of Bladder Organoids into Recipient Murine Bladder

Abstract

Source: Kim, Y. et al. Culture, Manipulation, and Orthotopic Transplantation of Mouse Bladder Tumor Organoids. J. Vis. Exp. (2020)

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.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Subculture Bladder Tumor Organoids

NOTE: Passage of bladder tumor organoids when they reach 100–150 µm in diameter is recommended.

1. Add 500 µL of collagenase/dispase to the organoid medium in a 24-well plate with tumor organoids. Pipette up and down the basement membrane matrix and the medium. Incubate for 20 min at 37 °C and harvest the cells into a 15 mL tube.
   NOTE: Examine....

Results

Figure 1: Orthotopic transplantation of bladder tumor organoids. (A) Schematic diagram of orthotopic transplantation of bladder tumor organoids to a nude mouse. (B) Representative images of bladders and.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
15 mL conical tube	 SPL	 50015
24-well plate	 Corning	 3526
29 G 1/2 insulin syringe	 SHINA	 B299473538
50 mL conical tube	 SPL	 50050
A8301	 Tocris	 2939	 Stock concentration: 25 mM
Absolute ethanol	 Daejung	 4023-2304
Absorbable suture	 Henry Schein	 039010
Advanced DMEM/F-12	 Thermo	 12634028
CAnN.Cg-Foxn1nu/Crl (nude mouse)	 Charles River	 194
Collagenase type I	 Thermo	 17100017	 Stock concentration: 20 mg/mL
Collagenase type II	 Thermo	 17100015	 Stock concentration: 20 mg/mL
Collagenase/dispase	 Sigma	 10269638001	 Stock concentration: 1 mg/mL
DMEM (Dulbecco's modified minimum essential media)	 Gibco	 11965-118
DMSO (Dimethyl sulfoxide)	 Sigma	 D8418
DPBS (Dulbecco's phosphate-buffered saline)	 Welgene	 LB 001-02
Matrigel growth factor reduced (GFR) Growth Factor Reduced (GFR)	 Corning	 354230	 Use for organoid culture in plate
Matrigel high concentration (HC)	 Corning	 354248	 Use for organoid transplantation
1.5 mL microtube	 Axygen	 MCT-150-C
Razor blade
Saline buffer	 JW Pharmaceutical
Trypsin-EDTA (0.25%)	 Gibco	 25200072
Ultracentrifugation tube	 Beckman Coulter	 331372
Y-27632 dihydrochloride	 Abmole	 M1817	 Stock concentration: 10 mM