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生長狀態(tài)：貼壁生長
年限：48 years
ATCC Number：CRL-2128?
相關(guān)疾?。耗[瘤
細(xì)胞形態(tài)：上皮樣
是否是腫瘤細(xì)胞：1
物種來源：人
運輸方式：凍存運輸
器官來源：腎上腺
組織來源：cortex
數(shù)量：大量
規(guī)格：0.1ml Designations: NCI-H295R [H295R]
Depositors: ?WE Rainey
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Homo sapiens
Morphology:epithelial


Source: Organ: adrenal gland
Tissue: cortex
Disease: carcinoma
Cellular Products:aldosterone; cortisol; C19 steroids
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Applications:It is responsive to angiotensin II and potassium ions.
NCI-H295R was adapted from the NCI-H295 pluripotent adrenocortical carcinoma cell line (ATCC CRL-10296 ) established by A.F. Gazdar and associates from a carcinoma of the adrenal cortex.
This cell line retains the ability to produce adrenal androgens.
DNA Profile (STR):Amelogenin: X
CSF1PO: 10,12
D13S317: 13
D16S539: 11
D5S818: 12
D7S820: 9,12
THO1: 9.3
TPOX: 8
vWA: 17,18
Age: 48 years
Gender: female
Ethnicity: Black
Comments:NCI-H295R was adapted from the NCI-H295 pluripotent adrenocortical carcinoma cell line (ATCC 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. This cell line retains the ability to produce adrenal androgens. It is responsive to angiotensin II and potassium ions.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No. 30-2006. To make the complete growth medium, add the following components to the base medium: 0.00625 mg/ml insulin; 0.00625 mg/ml transferrin; 6.25 ng/ml selenium; 1.25 mg/ml bovine serum albumin; 0.00535 mg/ml linoleic acid; adjust to a final concentration of 2.5% Nu-Serum I. The additives (insulin, transferrin, selenium, BSA and linoleic acid) are available in the form of ITS+ Premix from BD Biosciences. Nu-Serum is also available from BD Biosciences.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Subculturing: Protocol:

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which 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 37C to facilitate dispersal.
4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.
6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
7. Incubate cultures at 37?C.
   Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
   Medium Renewal: 2 to 3 times per week

Preservation: Freeze medium: Complete growth medium + extra 7.5% Nu-Serum, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Related Products:Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2006
parental cell line:ATCC CRL-10296
0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
References: 22400: Rainey WE, et al. The NCI-H295 cell line: a pluripotent model for human adrenocortical studies. Mol. Cell. Endocrinol. 100: 45-50, 1994. PubMed: 8056157
22764: Rainey WE, et al. Regulation of human adrenal carcinoma cell (NCI-H295) production of C19 steroids. J. Clin. Endocrinol. Metab. 77: 731-737, 1993. PubMed: 8396576
23082: Gazdar AF, et al. Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis. Cancer Res. 50: 5488-5496, 1990. PubMed: 2386954
23515: Bird IM, et al. Human NCI-H295 adrenocortical carcinoma cells: a model for angiotensin- II-responsive aldosterone secretion. Endocrinology 133: 1555-1561, 1993. PubMed: 8404594