Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

The Renca cell line was derived from a tumor that arose spontaneously as a renal cortical adenocarcinoma in Balb/cCr mice.

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The pattern of growth of this tumor accurately mimics that of human adult renal cell carcinoma, particularly with regard to spontaneous metastasis to lung and liver.xa0RefMurphy GP, Hrushesky WJ. A murine renal cell carcinoma. J. Natl. Cancer Inst. 50(4):1013-25, 1973. PubMed: 4703766xa0RefSalup RR, et al. Role of natural killer activity in development of spontaneous metastases in murine renal cancer. J. Urol. 134(6):1236-41, 1985. PubMed: 4057425xa0The cells do not express transforming growth factor-beta type II receptor (TbetaR-II) 10414746.xa0RefEngel J, et al. Transforming growth factor-beta type II receptor confers tumor suppressor activity in murine renal carcinoma (renca) cells . Urology. 54(1):164-70, 1999. PubMed: 10414746

Volumes used in this protocol are for 75 cm² flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbeccos phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 2 X 10⁴ to 4 X 10⁴ viable cells/cm² is recommended.
6. Incubate cultures at 37°C. We recommend that you subculture when the culture reaches a cell concentration between 8 X 10⁴ and 1.5 X 10⁵ cells/cm².

Medium renewal: Every 2 to 3 days.

Freeze medium: RPMI-1640 Medium, 77.5%; FBS, 15% FBS; DMSO, 7.5%

Storage temperature liquid nitrogen vapor phase

Temperature: 37°C

Atmosphere: air, 95%; carbon dioxide (CO₂), 5%