HAC15 cell line人肾上腺皮质癌细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

HAC15 cell line人肾上腺皮质癌细胞株

Product category
Human cells
Organism
Homo sapiens, human
Morphology
epithelial-like
Tissue
Adrenal gland
Disease
Carcinoma
Applications
3D cell culture

Specific applications
HAC15 provides an important model system for defining the molecular mechanisms regulating aldosterone and cortisol production and can be applied to studies of either normal adrenal cell function or adrenocortical cancer.

Characteristics
Growth properties
Adherent
Derivation
HAC15 was clonally isolated from NCI-H295R (BioVector® CRL-2128™). NCI-H295R was adapted from the NCI-H295 pluripotent adrenocortical carcinoma cell line (BioVector® CRL-10296™) established by A.F. Gazdar and associates from a carcinoma of the adrenal cortex. The original cells were adapted to a culture medium which decreased the population doubling time from 5 days to 2 days. While the original cells grew in suspension, the adapted cells were selected to grow in a monolayer. The HAC15 cell line responds to ACTH, whereas NCI-H295R lacks this responsiveness.
Age
48 years
Ethnicity
Black
Gender
Female
Genes expressed
aldosterone; cortisol; C19 steroids
Comments
HAC15 is a clonal cell line isolated from NCI-H295R which, in extended culture, displays a more stable steroidogenic phenotype than NCI-H295R. The HAC15 cells increase aldosterone production in response to treatment with angiotensin II and high extracellular potassium levels. These cells respond to activation cAMP signaling pathway agonists, forskolin and dibutyryl-cAMP, with a time-dependent increase in cortisol and dehydroepiandrosterone production. HAC15 also exhibit a modest increase in cortisol following chronic ACTH (adrenocorticotropic hormone) treatment. In summary, the HAC15 cell line is capable of responding to the three main adrenocortical physiologic regulators.
Handling information
Unpacking and storage instructions
1.Check all containers for leakage or breakage.
2.Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below _x001f_-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
To make complete growth medium add to 500 mL DMEM: F12 Medium (BioVector® 30-2006™): 5.6 mL ITS + Premix (Becton Dickenson Cat. No. 354352) and 56 mL Cosmic Calf Serum (Hyclone Cat. No. SH30087.03)
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure
To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.  
1.Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
2.Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3.Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.
4.Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).
5.Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

Subculturing procedure
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
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 that contains trypsin inhibitor.
3.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.
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. An inoculum of 3 X 104 to 6 X 104 viable cells/cm2 is recommended.
6.Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 2 X 105 and 3 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week

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