Modeling Metastatic Endometrial Cancer Model: A Technique to Generate Endometrial Cancer with Lymph Node Metastasis in Rabbit Models

Abstract

Source: Lauren Philp et al. An Orthotopic Endometrial Cancer Model with Retroperitoneal Lymphadenopathy Made From In Vivo Propagated and Cultured VX2 Cells. J. Vis. Exp. (2019)

This video demonstrates the generation of a rabbit model of endometrial cancer by injecting a highly metastatic VX2 cell suspension into the sutured uterine horns of the rabbit. This leads to generation of primary endometrial cancer with retroperitoneal lymph node metastases.

Protocol

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

1. Creation of in vitro VX2 cell line

1. Harvest the VX2 tumor from rabbit quadricep muscle and growth in mouse flank.
   1. Thaw frozen VX2 tumor blocks (1 cm x 1 cm), mince on a culture plate using a scalpel blade and strain through a 70 µm filter by adding small amounts of Hanks Balanced Salt Solution (HBSS) sequentially to ensure all cells are strained for a final volume of 1 mL.
   2. Count cells and dilute with 0.9% phosphate buffered....

Results

Figure 1: Uterine suturing. Black arrow = sutures, red arrow = uterine horns.

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Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
11-blade scalpel, Sterile, Disposible	Aspen Surgical (VWR)	80094-086
22-gague ear vein catheter	CDMV	14332
3-0 absorbable poly-filament suture (Polysorb)	Covidien	356718
3-0 braided absorbable suture (Polysorb)	Covidien	356718
70uM cell strainer, Individually wrapped, Nylon	Falcon	352350
Acepromazine (Atravet)	CDMV	1047
Betadine soap (Poviodone iodine 7.5%)	CDMV	4363
Betadine solution (Poviodone iodine 10%)	UHN Stores	457955
Buprenorphine	McGill University
Cefazolin	UHN in-patient pharmacy	No Cat # Needed
Chlorhexidine solution	CDMV	119872
Corning BioCoatCellware, Collagen Type I, 100mm dishes	Corning	354450	Brand not important
Corning BioCoatCellware, Collagen Type I, 24-well plates	Corning	354408	Brand not important
Corning BioCoatCellware, Collagen Type I, 6-well plates	Corning	354400	Brand not important
Corning Matrigel Basement Membrane Matrix, *LDEV-free, 10 mL	Corning	354234
DMEM/HAM F12 1:1	Life Technologies	11320	Brand not important
DMSO	Caledon Lab Chem	10/1/4100
Enrofloxacin (Baytril injectable)	CDMV	11242
Falcon Tube	Corning Centri-Star	430828
Fetal Bovine Serum, Qualified, Canadian Origin, 500ml	Life Technologies	12483020	Brand/source not important
Isoflurane	UHN in-patient pharmacy	No Cat # Needed
Isohexol contrast	GE Healthcare	407141210
Meloxicam (Metacam 0.5%)	CDMV	104674
Normal Saline	House Brand (UofT Medstore)	1011
PBS	Multicell or Sigma	331-010-CL or D8537-500mL
Penicillin/Streptomycin (100mL; 10000U Penicillin, 10000ug Streptomycin)	Corning-Cellgro	CA45000-652
Sterile Hanks Balanced Salt Solution (-Ca++, -Mg++, -Phenol Red)	T.C.M.F (Dr Bristow)	28-Jan-11
Surgical Glue (Tissue Adhesive)	3M Vetbond	14695B
Trypsin (0.25%), Proteomics Grade	Sigma	T-6567-5X20UG
Trypsin-EDTA, 0.05%, 100ml	Wisent Inc	325-542-EL	Brand not important