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

プロトコル

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 hemocytome....

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資料

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