PC-12 Cell Line from rat

Rat adrenal phaeochromocytoma

Derived from a transplantable rat adrenal phaeochromocytoma. Respond reversibly to nerve growth factor by induction of neuronal phenotype. Cells synthesise and store catecholamine, dopamine and NPP. Used in neurobiological and neurochemical studies. When cells are grown with collagen - neuronal fibroblasts; without collagen - spherical clusters. PLEASE NOTE: Growing orders of PC-12 despatched from ECACC are grown without collagen and therefore the cells will be in suspension on receipt.

Neurobiological and neurochemical studies

PC-12 cell line from rat has been used to create artificial nervous system tissue models to study neurodegeneration. It has also been used to perform cell contact assay to test the biocompatibility of high-density electrode arrays.

RPMI 1640 + 2mM Glutamine + 10% Horse Serum + 5% Foetal Bovine Serum (FBS)

Cells grow in small clusters but adhere poorly to plastic. The cells do grow satisfactorily in suspension in untreated flasks. Maintain cultures between 2-5x100,000 cells/ml: 5% CO₂; 37°C. For attachment grow in collagen coated flasks (must be collagen type IV), feed 3 times a week and split cultures 1:3 to 1:6 (i.e. seeding at 2-4 x 10,000 cells / cm²) using 0.25% trypsin/EDTA; 5% CO₂; 37°C. Growing cultures will be supplied in suspension. On receipt, incubate the flask overnight without opening and on introduction to a type IV collagen coated flask the cells should attach. Preparation of collagen solution: Use collagen type IV (Sigma C5533). Add 5mg collagen to 50ml 0.1M glacial acetic acid to obtain a 0.01% collagen solution. Stir at room temperature for 1-3 hours. Sterilise by transferring the solution to a screw capped glass bottle and carefully adding an aliquot of chloroform which will form a layer beneath the collagen solution. The volume of chloroform should be approximately 10% that of collagen. After overnight incubation at 4C aseptically remove the top layer of collagen and store in sterile tubes or flasks at 4oC. NB: Growth factor reduced matrigel may also be used. Preparation of flasks: Add sufficient collagen to cover the surface of the required number of tissue culture flasks and leave for at least 6 hours at 37°C. Remove excess fluid and allow flasks to dry by incubating at 37°C, leaving the caps loose. Prior to addition of cells wash flask three times in PBS. Flasks can be purchased from suppliers pre-coated with type IV collagen. Reported doubling time = 92 hours

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