Subcutaneous Transplantation Assay: A Method to Transplant Melanoma Cells from a Donor to a Recipient Zebrafish Model
Published: April 30, 2023
Abstract
Source: Iyengar, S. et al. Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model. J. Vis. Exp.(2012)
This video describes the method for transplantation of tumor cells from the melanoma-bearing donor zebrafish model to an immunocompromised recipient Casper zebrafish. It helps to visualize tumor engraftment and proliferation in the recipient fish.
Protocol
All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Transplantation Assay Prepare recipient 2-3 month old Casper fish by irradiating with 25 Gy of gamma irradiation one day prior to transplantation. Allow the fish to recover in fish water. Euthanize a tumor-bearing fish according to institutional guidelines. Specifically, a fish is placed into a dish with 0….
Representative Results
Figure 1. Transplantation of melanoma cells. A) Uninjected Casper zebrafish. Scale bar = 500 μM. B) Subcutaneous transplantation site (arrowhead) on an irradiated Casper recipient immediately after injection with 50,000 melanoma cells. Scale bar = 200 μM. C) Irradiated Casper recipient showing…
開示
The authors have nothing to disclose.
Materials
| Gateway recombination reagents | Invitrogen | ||
| miniCoopR | |||
| casper Zebrafish | |||
| 701N 10 μl Syringe | Hamilton/Fisher | 14-824 | |
| 40 μM filter | BD Falcon/Fisher | 352340 | |
| FBS | Invitrogen | 26140079 |
