Long-term Orthotopic Transplantation of Tumor Cells into Zebrafish Embryos: An Efficient Method to Deliver Tumor Cells into the Fourth Ventricle of Zebrafish Embryos for Long-term Analysis of Tumor Cell Behavior
Published: April 30, 2023
Abstract
Source: Casey, M. J. et al. Transplantation of Zebrafish Pediatric Brain Tumors into Immune-competent Hosts for Long-term Study of Tumor Cell Behavior and Drug Response. J. Vis. Exp. (2017)
This video demonstrates the orthotopic transplantation of zebrafish brain tumor cells into the fourth ventricle of an immunocompetent zebrafish embryo. This procedure allows for the long-term study of tumor cell invasion and behavior.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Injecting the Tumor Suspension into the Fourth Ventricle of a 2-dpf Embryo Transfer 10–20 anesthetized dechorionated embryos to the periphery of the injection plate using a transfer pipette; the embryos should fall laterally onto the injection plate, with the ventricles clearly visible and accessible. Use an angle…
Divulgaciones
The authors have nothing to disclose.
Materials
| Egg water | In house | Maintaining embryos, making injection plate | |
| Petri dish | Thermo Fisher | FB0875711Z | Housing embryos, making injection plate |
| Glass Container | Any commercial brand | Making injection plate | |
| Micropipettes | Any commercial brand | Tumor resuspension and transplantation | |
| Angled Probe | Fine Science Tools | 10140-02 | Embryo manipulation |
| Transfer Pipette | Any commercial brand | Embryo manipulation | |
| Stereomicroscope | Olympus | SZ61 | Tumor transplantation |
| Fluorescent Stereomicroscope | Olympus | SZX16 | Imaging tumor transplants |
| Microscope Camera | Olympus | DP-72 | Imaging tumor transplants |
| Incubator | Any commercial brand | Maintaining embryos, warming up injection plate | |
| N2 | Any commericial brand | For microinjector set up | |
| Glass capillary (no filament) | World Precision Instruments | TW120-4 | Tumor transplantation |
| Needle puller | Sutter Instruments | P-97 | Tumor transplantation |
| Microloader tips | Eppendorf | 930001007 | Tumor transplantation |
| Microinjector | Harvard Apparatus | PLI-90 | Tumor transplantation |
| Tricaine-S (MS-222) | Western Chemical | TRS1 | Tumor transplantation, anesthetic |
| Agarose | Denville | CA3510-8 | Making injection plate |
| Microwave | Any commercial brand | Making injection plate | |
| Scale | Any commercial brand | Making injection plate | |
| Gloves | Any commercial brand | All aspects of the protocol | |
| Pronase | Roche Diagnostics | 11459643001 | Dechorionate embryos |
| Razor Blade | Thermo Fisher | 12640 | Needle preparation |
| 50 mL beaker (2 inch diameter) | Any commercial brand | Making injection plate | |
| Glass Container | Any commercial brand | Making injection plate | |
| Methylene Blue | Sigma-Aldrich | M9140 | Add to egg water to prevent fungal growth |
Tags
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Long-term Orthotopic Transplantation of Tumor Cells into Zebrafish Embryos: An Efficient Method to Deliver Tumor Cells into the Fourth Ventricle of Zebrafish Embryos for Long-term Analysis of Tumor Cell Behavior. J. Vis. Exp. (Pending Publication), e20689, doi: (2023).