The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a “patient-like” orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors.
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Method Article
A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis
In This Article
Summary
Abstract
The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of cancer metastasis are still unclear. Animal models are essential for elucidating the mechanisms and for evaluating novel strategies for the treatment of metastatic cancers. Here, an in-depth description of a “patient-like” orthotopic syngeneic mouse model for exploring the mechanisms of metastasis of solid organ tumors is provided. The survival surgical implantation of BNL 1ME A.7R.1 mouse hepatocellular carcinoma cells directly into the liver (the organ of origin) of the inbred wild-type immune competent laboratory mouse strain, BALB/c is described. The success and reproducibility of this methodology recommends it for widespread use in elucidating the biological mechanisms of solid organ cancer metastasis.
Introduction
Hepatocellular carcinoma (HCC) is one of the most lethal cancers, with poor prognosis and low life expectancy. Nearly all cancer-related deaths are due to the metastatic spread of the disease from the originating organ to additional distant organs2-6.HCC progression is a complex process. Therefore, being able to model the tumor microenvironment that is naturally found in metastatic HCC in animal models can prove to be a successful and useful way to reveal relevant mechanisms in humans.7Unfortunately, the mechanisms of cancer metastasis are still largely unclear. Therefore, there is a need to establish animal models that will enable us to elucidat....
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Protocol
Ethics Statement
All animal studies were approved by the Institutional Animal Care and Use Committee (IACUC) of Hunter College of The City University of New York.
Note: Eight BALB/c mice were used in this experimental procedure. Five BALB/c mice were implanted with 5 X 106 BNL 1ME A.7R.1 mouse hepatocellular carcinoma cells to produce the primary tumors. Three BALB/c mice with no implantations were used as controls. BNL 1ME A.7R.1 is a murine hepatocellular carcinoma cell line that was derived by chemical transformation from the murine non-tumorigenic hepatocyte cell line BNL CL.2. BNL 1ME A.7R.1 was obtain....
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Results
The livers of Balb/c mice were implanted with 5×106 BNL 1ME A.7R.1 mouse HCC cells. Clinical evidence of HCC development was observable 63 days post-surgery. Consequently, mice were humanely euthanized. Necropsy was performed on the euthanized mice. The lungs and liver were resected and a careful gross examination was performed to identify macroscopic tumors. In one mouse, a superficial tumor was observed on the surface of the liver with some attachment to the abdominal wall (Figure 1A). In a .......
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Discussion
In this article an in-depth description of the method is given that was recently reported by Ogunwobi and colleagues of successful establishment of an orthotopic syngeneic mouse model of HCC metastasis (Figure 2)3. The take rate of tumors for this procedure is generally high. We have previously observed a tumor take rate of 100%3. However, the take rate can be variable depending on the competence of the investigator. In a recent experiment performed by a new graduate student under t.......
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Disclosures
The authors declare that they have no competing financial interests.
Acknowledgements
We would like to thank Pascal DuBois for proof-reading the article. The authors would also like to thank the personnel of the Animal Facility at Hunter College of The City University of New York. This work was supported by a NIH RCMI grant to Hunter College.
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Micro Dissecting Tweezer | Roboz | (RS-5040) | Tip 0.20 x 0.12 mm |
| Graefe Micro Dissecting Forceps, serrated curved tip | Roboz | (RS-5111) | 1 X 2 teeth, curved tip width 0.6 mm |
| Micro Dissecting Retractor-Agricola (3 by 3 prongs) | Roboz | (RS-6501) | Blunt 3 X 3 prongs, depth 4 mm, spread 25 mm |
| Micro Dissecting Retractor-Goldstein (3 by 3 prongs) | Roboz | (RS-6503) | Blunt, 3 X 3 prongs, depth 4 mm, spread 19 mm |
| Jameson Caliper (Measures tumors) | Roboz | (RS-6466) | 80 mm/3 inch scale, chrome plated |
| Micro Dissecting Scissors, large ring sissors, straight and sharp | Roboz | (RS-5852) | 23 mm blades, length 4 inches, flat shanks and large rings |
| Scalpel with blades, for delicate dissecting procedures | Roboz | (RS-9861-36) | |
| Scalpel handle | Roboz | (RS-9884) | Solid |
| Littauer Stitch Sissors | Roboz | (RS-7074) | Length 4.5 inches |
| Brown needle holder, for easy suture tying | Roboz | (RS-7960) | Convex jaw, fine serrations |
| Reflex 7MM wound clips with reflex 7 clip applier | Roboz | (RS-9262) | safe, secure alternative method of wound closure |
| Instrument tray and lid | Roboz | RT-1350S | |
| Mini-clipper with detachable blade | Roboz | (RS-5903) | |
| Germinator 500 (the Germ Terminator) Dry Sterilizer | Roboz | Ds-400, Ds-401, DS-501 | For fast decontamination of micro-dissecting instruments. Instruments decontaminate within 15 sec. |
| Microinjection needles | VWR | BD305125 | 25G Needle |
References
- Sheng, W., et al. Capture, release and culture of circulating tumor cells from pancreatic cancer patients using an enhanced mixing chip. Lab on a Chip. 14, 89-98 (2014).
- Ogunwobi, O. O., Liu, C.
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