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

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記事を引用
Zebrafish Xenograft Assay: A Method to Assess the Anti-cancer Efficacy of Therapeutics on Tumorigenicity in Zebrafish In Vivo. J. Vis. Exp. (Pending Publication), e20244, doi: (2023).

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