| Field | Specification |
|---|---|
| Species |
NCI-H1299, also known as H1299, is a cell line established from a lymph node metastasis of the lung from a 43-year-old white male patient with carcinoma. H1299 and H292 are non-small cell lung cancer (NSCLC) cell lines.
Regarding their genetic profile, H1299 cells have a homozygous partial deletion of the p53 protein and lack expression of p53 protein. While KRAS mutations are commonly found in various types of cancer, including NSCLC, H1299 expresses KRAS WT. A549 is another NSCLC cell line that homozygously expresses endogenous KRAS G12S.
Understanding the biology of KRAS and its downstream signalling pathways is crucial for developing effective cancer therapies. Therefore, this epithelial-like cell line is commonly used in cancer and immuno-oncology research.
The morphology of H1299 cells is characterized by adherent flattened cells with a thickness of fewer than 5 microns. H1299 cells have an approximate doubling time of 22 - 30 hours. H1299 cells express keratin and vimentin but are negative for neurofilament triplet protein.
They are also reported to be able to synthesize the peptide neuromedin B (NMB) at 0.1 pmol/mg protein but not the gastrin-releasing peptide (GRP). Compared to A549 cells with more epithelial characteristics, H1299 cells have more mesenchymal characteristics and less effective epithelial marker expression.
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- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS, add 2.5 g/L glucose and 10 mM HEPES
- 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
- 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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