NCI-H295R細(xì)胞

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

年限： 48 years

ATCC Number： CRL-2128?

相關(guān)**： 腫瘤

細(xì)胞形態(tài)： 上皮樣

是否是腫瘤細(xì)胞： 1

物種來(lái)源： 人

運(yùn)輸方式： 凍存運(yùn)輸

器官來(lái)源： 腎上腺

組織來(lái)源： cortex

數(shù)量： 大量

NCI-H295R細(xì)胞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

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Applications: NCI-H295R細(xì)胞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細(xì)胞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:

Remove and discard culture medium.

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.

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.

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.

Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.

Incubate cultures at 37?C.

Subcultivation Ratio: NCI-H295R細(xì)胞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