PC-12 cells

Disease:

Rat adrenal pheochromocytoma

Origin:

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: PC-12 cells from us are grown without collagen and therefore the cells will be in suspension on receipt.

Species:

Rat, Rattus norvegicus

Tissue:

Adrenal gland

Morphology:

small irregularly shaped cells

Properties:

Suspension, floating clusters; few scattered loosely attached cells

Cytogenic data:

40 chromosomes; 38 autosomes plus XY

Patient:

Male

Medium:

RPMI-1640 + 2 mM Glutamine + 10% Horse Serum + 5% FBS

Subculture:

The cells do grow satisfactorily in suspension in untreated flasks. Maintain cultures between 2-5x100, 000 cells/ml. 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. Culture at 5% CO2, 37C.

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. Store in sterile glass bottles or as pre-coated flasks at 4C. 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 37C. Remove excess fluid and allow flasks to dry by incubating at 37C, 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

Freezing Medium:

Complete culture medium supplemented with 5% (v/v) DMSO