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NCI-H295R [H295R]
NCI-H295R [H295R]
規格:
價格:
編號:B167201
品牌:Mingzhoubio

標準菌株
定量菌液
DNA
RNA

規格:
凍干粉
斜面
甘油
平板


產品名稱
NCI-H295R [H295R]
商品貨號
B167201
Organism
Homo sapiens, human
Tissue
adrenal gland/cortex
培養基
Product Format
frozen
Morphology
epithelial
Culture Properties
adherent
Biosafety Level
1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease
Carcinoma
Age
48 years
Gender
female
Ethnicity
Black
Applications
This line has the ability to produce adrenal androgens.
Storage Conditions
liquid nitrogen vapor phase
Images
Derivation
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.
Clinical Data
48 years
Black
female
Genes Expressed
aldosterone; cortisol; C19 steroids
Cellular Products
aldosterone; cortisol; C19 steroids
Comments
It is responsive to angiotensin II and potassium ions.
This cell line retains the ability to produce adrenal androgens.
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 Corning (Catalog No. 354352). Nu-Serum is also available from Corning (Catalog No. 355100).
Subculturing
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  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 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. 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 cell suspension to new culture vessels.
  7. Incubate cultures at 37°C.

Subculture Ratio: 1:3 to 1:4
Medium Renewal: 2 to 3 times a week.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 4th edition, published by Wiley-Liss, N.Y., 2004.

Cryopreservation
Freeze medium: Complete growth medium + extra 7.5% Nu-Serum, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X
CSF1PO: 10,12
D13S317: 13
D16S539: 11
D5S818: 12
D7S820: 9,12
THO1: 9.3
TPOX: 8
vWA: 17,18
Name of Depositor
WE Rainey
Deposited As
Homo sapiens
References

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

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

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

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

Rainey WE, et al. Adrenocortical cell lines. Mol. Cell Endocrinol. 228(1-2): 23-38, 2004. PubMed: 15541570

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