| Field | Specification |
|---|---|
| Species |
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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Products: Aldosterone, cortisol, C19 steroids
- cultureMedium: You can purchase our ready-to-use NCI-H295R Cell Growth Medium (820402) or choose to supplement DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a) with the below additives
- 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
- dissociationReagent: 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.
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: 48 hours
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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