Orthotopic Injection: Implanting Tissue Specific Cancer Cells into an Adult Mouse

Abstract

Source: Katsuta, E., et al. Generating a Murine Orthotopic Metastatic Breast Cancer Model and Performing Murine Radical Mastectomy. J. Vis. Exp. (2018).

The video demonstrates the breast cancer orthotopic inoculation technique in a mouse model. This technique produces a high tumorigenesis rate with less variability in tumor size and shape compared to subcutaneous or non-surgical injection.

Protocol

1. Preparation of Cells (for 10 Mice)

NOTE: The cells should be inoculated within 1 h after being detached from the dish to avoid decreased cell viability. Specifically, the cell suspension should be mixed into the gelatinous protein mixture within 15 min after detaching the cells from the dish to maintain their viability.

1. Culture 4T1-luc2 cells, a mouse mammary adenocarcinoma cell line expressing luciferase, in RPMI 1640 media with 10% FBS in a humidified incubator at 37 °C in 5% CO₂.
2. Wash the adherent 4T1-luc2 cells in a 10-cm dish with phosphate-buffered saline (PBS) using a 10 ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Micro Dissection Scissors	Roboz	RS-5983	For cancer cell innoculation and mastectomy
5-0 silk sutures	Look	774B F
Roswell Park Memorial Insitute 1640	Gibco	11875093	For cell culture
Fetal Bovine Serum	Gibco	10437028	For cell culture
Trypsin-EDTA (0.25%)	Gibco	25200056	For cell culture
Dry sterilant (Germinator 500)	Braintree Scientific	GER 5287-120V	For cancer cell innoculation and mastectomy
Needle Holder	Roboz	RS-7830	For cancer cell innoculation and mastectomy
Adson Forceps	Roboz	RS-5233	For cancer cell innoculation and mastectomy
Clipper	Wahl	9908-717	For cancer cell innoculation and mastectomy
Matrigel	Corning	354234	For cancer cell innoculation