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Comments: The RIN-m5F cell line was is a clone derived from the RIN-m rat islet cell line. The cells produce and secrete insulin, and produce L-dopa-decarboxylase a marker for cells havingamine precursor uptake and decarboxylation or APUD, activity . Unlike the parental line they do not produce somatostatin. Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.Temperature: 37.0°C Subculturing: Protocol: Remove and discard culture medium. Briefly rinse the cell layer with 0.25% w/v Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. 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. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Cells will take about 3 days to attach fully. Cells attach as small islands which begin to flatten and spread into patchy epithelial morphology by day 3 or 4. Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended Medium Renewal: Two to three times weekly. Do not fluid change for 3 to 4 days after plating. Cells are loose. Just add medium. Preservation: Freeze medium: Complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor phase Modified to contain 2 mM L-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 4500 mg/L glucose, and 1500 mg/L sodium bicarbonate.NOTE: This reduced level of