Generating Renal Cell Carcinoma Model: A Protocol to Develop Orthotopic RCC Murine Model by Intrarenal Implantation of Cancer Cells

Abstract

Source: Murphy, K. A. et al. A Syngeneic Mouse Model of Metastatic Renal Cell Carcinoma for Quantitative and Longitudinal Assessment of Preclinical Therapies. J. Vis. Exp. (2017)

In this video, we develop an orthotopic mouse model of renal cell carcinoma. This model results in reproducible primary tumors in the injected kidney and metastatic tumors in the lung, allowing the evaluation of disease progression and treatment.

Protocol

1. Implantation of Cells

To implant Renca cells intrarenally, prepare Renca cells into a single-cell suspension at 2 x 10⁶ cells/mL in Hank's Balanced Salt Solution (HBSS).

1. Remove adherent Renca cells from the tissue culture flask using 0.25% trypsin in HBSS. After ~5 min, add complete RPMI to the flask to neutralize the trypsin and transfer the cells to a centrifuge tube for washing. Centrifuge the cells at 300 relative centrifugal g force (rcf x g) for 5 min at 25 °C.
2. Decant the supernatant and resuspend the cells in HBSS. Count the cells using a hemocytometer ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Artificial tears ophthalmic ointment	Akorn	NDC 17478-162-35
Vetbond Tissue Adhesive	3M	CBGBIW011019
Betadine Solution (Povidoneiodine, 5%)  NDC	Purdue Products, L.P.	67618-155-32
0.25% Marcaine (Bupivavaine HCl injection)	Hospira	0409-1587-50
Heating pad	Sunbeam
Syringes BD		309659
Gauze	Venture	908291
Ketamine	Phoenix Pharmaceuticals NDC	57319-609-02
Xylazine	Akorn NDC	59399-111
Hank’s Balanced Salts	Sigma-Aldrich	H2387
Scissors
Forceps
Renca	ATCC	CRL-2947