Zebrafish Xenograft Assay: A Method to Assess the Anti-cancer Efficacy of Therapeutics on Tumorigenicity in Zebrafish In Vivo
Published: April 30, 2023
Abstract
Source: Lee, J. Y et al.,Preclinical Assessment of the Bioactivity of the Anticancer Coumarin OT48 by Spheroids, Colony Formation Assays, and Zebrafish Xenografts. J. Vis. Exp. (2018)
This video describes the zebrafish xenograft assay to assess in vivo tumor formation and study the effect of therapeutic agents.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Zebrafish Xenograft Assay NOTE: This technical approach is visualized as a schematic (Figure 1). Preparation of stock reagents Prepare 1 L of 30x Danieau’s solution: 1740 mM NaCl, 21 mM KCl, 12 mM MgSO4•7H2O, 18 mM Ca(NO<sub…
Representative Results
Figure 1: Schematic diagram of the overall process of the zebrafish xenograft assay.
開示
The authors have nothing to disclose.
Materials
| A549 | ATCC | CCL-185 | 37 C° |
| RPMI 1640 | Lonza | 30096 | 4 C° |
| FBS | Biowest | S1520-500 | -20 C° |
| Penicillin-streptomycin | Lonza | 17-602E | -20 C° |
| Cell culture flask T25 | SPL | 70025 | RT |
| Cell culture flask T75 | SPL | 70075 | RT |
| Cell culture flask T175 | SPL | 71175 | RT |
| 1.5ml tube | Extragene | Tube-170-C | RT |
| 24 well plate | SPL | 30024 | RT |
| Petridish | SPL | 10100 | RT |
| PBS solution | Hyclone | SH30256.02 | RT |
| Trypsin-EDTA | Gibco | 25-300-054 | 4 C° |
| Sodium Chloride | Sigma-Aldrich | 71382 | RT |
| Potassium chloride (KCL) | Sigma-Aldrich | P9541 | RT |
| Magnesium sulfate heptahydrate (MgSO4.7H2O) | Sigma-Aldrich | M2773 | RT |
| Calcium nitrate tetrahydrate (Ca(NO3)2) | Sigma-Aldrich | C1396 | RT |
| HEPES solution | Sigma-Aldrich | H0887 | RT |
| Ethyl 3-aminobenzoate methanesulfonate (Tricaine) | Sigma-Aldrich | E10521 | RT |
| Phenol Red solution | Sigma-Aldrich | P0290 | RT |
| Methylcellulose | Sigma-Aldrich | M0512 | RT |
| 1-phenyl-2-thiourea (PTU) | Sigma-Aldrich | P7629 | RT |
| CM-Dil dye | Invitrogen | C7001 | -20 C° |
| Glass capillary | World Precision Instruments | TW 100F-4 | RT |
| Micropipette puller | Shutter instrument, USA | P-97 | RT |
| Micro injector | World Precision Instruments | PV820 | RT |
| Syringe | KOVAX | 1ml | RT |
| Micro loader | Eppendorf | 5242956003 | RT |
| Glass slide | Marienfeld | HSU-1000612 | RT |
| Fluorescence microscopy | Leica | DE/DM 5000B | RT |
