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These cells offer models for the study of the biology of pancreatic islet cells, specifically the mechanisms controlling the synthesis, storage and secretion of insulin and somatostatin.

The RIN-5F cell line is a clone derived from the RIN-m rat islet line (ATCC CRL-2057 ). The cells produce and secrete insulin, and produce L-dopa-decarboxylase (a marker for cells having amine precursor uptake and decarboxylation, or APUD, activity). Unlike the parental line they do not produce somatostatin. The parental line was also cloned to establish RIN-14B (ATCC CRL-2059) that produces only somatostatin. -

male

insulin; L-dopa-decarboxylase

insulin; L-dopa-decarboxylase

Note:  Monolayer never becomes confluent.

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 Dulbecco's 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 1.0 to 2.0 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.
6. Incubate cultures at 37°C.

Note:  Cells will take about 3 days to attach fully.

Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended

Medium Renewal: Every 3 to 4 days

Freeze medium: Complete growth medium, 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor temperature

Temperature: 37°C