NCI-H295R Cells
General information
Description | H295R was adapted from the NCI-H295 pluripotent adrenocortical carcinoma cell line established by A.F. Gazdar and associates (1990) 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. The adapted cells were selected to grow in a monolayer, in contrast to the original cells which grew in suspension. This cell line retains the ability to produce adrenal androgens. It is responsive to angiotensin II and potassium ions. |
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Organism | Human |
Tissue | Adrenal gland |
Disease | Carcinoma |
Synonyms | NCI-H295R, NCI H295R, NCIH295R, H-295R, H295R-S1 |
Characteristics
Age | 48 years |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | Monolayer, adherent |
Identifiers / Biosafety / Citation
Citation | NCI-H295R (Cytion catalog number 300483) |
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Biosafety level | 1 |
Expression / Mutation
Products | Aldosterone, cortisol, C19 steroids |
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Handling
Culture Medium | You can purchase our ready-to-use NCI-H295R Cell Growth Medium (820402) or choose to supplement DMEM:Ham's F12, w: 3.1 g/L Glucose, w: 1.6 mM L-Glutamine, w: 15 mM HEPES, w: 1.0 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a) with the below additives |
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Medium supplements | Supplement the medium with 5% FBS, 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 |
Passaging solution | Accutase |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:3 to 1:4 is recommended |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | 48 hours |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,x
CSF1PO: 10,12
D13S317: 13
D16S539: 11
D5S818: 12
D7S820: 9,12
TH01: 9.3
TPOX: 8
vWA: 17,18
D3S1358: 15,16
D21S11: 32.2
D18S51: 17
Penta E: 5,12
Penta D: 8
D8S1179: 13
FGA: 19.2,24
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HLA alleles |
A*: 02:01:01
B*: 15:10:01
C*: 03:04:02
DRB1*: 01:01:01
DQA1*: 01:01:01
DQB1*: 05:01:01
DPB1*: 04:02:01
E: 01:03:02
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