| Comments |
The cells produce and secrete somatostatin, 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 insulin.
The parental line was also cloned to establish RIN-5F (ATCC CRL-2058) that produces only insulin. |
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM 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.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: Every 3 to 4 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| References |
Bhathena SJ, et al. Insulin, glucagon, and somatostatin secretion by cultured rat islet cell tumor and its clones. Proc. Soc. Exp. Biol. Med. 175: 35-38, 1984. PubMed: 6141566
Chick WL, et al. A transplantable insulinoma in the rat. Proc. Natl. Acad. Sci. USA 74: 628-632, 1977. PubMed: 191819
Bhathena SJ, et al. Insulin, glucagon, and somatostatin receptors on cultured cells and clones from rat islet cell tumor. Diabetes 31: 521-531, 1982. PubMed: 6295859
Gazdar AF, et al. Continuous, clonal, insulin- and somatostatin-secreting cell lines established from a transplantable rat islet cell tumor. Proc. Natl. Acad. Sci. USA 77: 3519-3523, 1980. PubMed: 6106192
Oie HK, et al. Clonal analysis of insulin and somatostatin secretion and L-dopa decarboxylase expression by a rat islet cell tumor. Endocrinology 112: 1070-1075, 1983. PubMed: 6129963
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
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