Orthotopic Inoculation of Thyroid Cancer Cells into Murine Thyroid: A Procedure to Establish Orthotopic Thyroid Cancer Mouse Model

Abstract

Source: Julia Ramirez-Moya, et al. In Vivo Inhibition of MicroRNA to Decrease Tumor Growth in Mice. J. Vis. Exp. (2019)

This video describes the procedure for orthotopic inoculation of thyroid cancer cells into the thyroid gland of mouse models, which may then serve as a valuable platform to assess carcinogenesis and discover potential treatment strategies.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Thyroid Orthotopic Inoculation of Cells

1. Cell preparation
   1. Engineer a Cal62 human thyroid cancer cell line to overexpress a transgenic construct that constitutively expresses GFP and luciferase. Select the transgenic cells by antibiotic resistance or by sorting the GFP-positive cells with flow cytometry.
   2. Grow these cells in DMEM supplemented with antibiotics (penicillin and streptomycin) and 10% FBS at 37 °C and 5% CO₂.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Basement Membrane Matrix: Matrigel Basement Membrane Matrix High Concentration	Corning	#354248
In vivo delivery reagent: Invivofectamine 3.0 Reagent	Thermo Fisher	IVF3005
In vivo imaging software: IVIS-Lumina II Imaging System	Caliper Life Sciences