Bladder Tumor Organoid Model: A Method for Orthotopic Transplantation of Bladder Organoids into Recipient Murine Bladder
Published: April 30, 2023
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. Add 500 µL of collagenase/dispase to the organoid medium in a 24-well plate with tumor organoids. Pipette up and down th…
Representative 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 H&E stained sections from mice orthotopically transplanted with bladder tumor organoids. Magnified views of the boxed regions in the mid…
Divulgaciones
The authors have nothing to disclose.
Materials
| 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 |
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Bladder Tumor Organoid Model: A Method for Orthotopic Transplantation of Bladder Organoids into Recipient Murine Bladder. J. Vis. Exp. (Pending Publication), e20553, doi: (2023).