NCI-H1299 Cells
Product number:
300485
Insights on NCI-H1299 lung cancer cells
Description | 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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Organism | Human |
Tissue | Lung |
Disease | Carcinoma |
Synonyms | H1299, H-1299, NCIH1299 |
Details
Age | 59 years |
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Ethnicity | Caucasian |
Growth properties | Adherent |
Characteristics of H1299 small cell lung cancer cells
Citation | NCI-H1299 (Cytion catalog number 300485) |
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Biosafety level | 1 |
Expression / Mutation
Handling
Culture Medium | RPMI 1640, w: 4.5 g/L Glucose, w: 2 mM L-Glutamine, w: 10 mM HEPES, w: 1 mM Sodium pyruvate, w: 1.5 g/L NaHCO3 (Cytion article number 820702a) |
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Medium supplements | Supplement the medium with 10% FBS |
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. |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures | NCI-H1299 cells are shipped in a deep-frozen state on dry ice. Upon receipt, confirm that the vial remains frozen. For storage, place the cryovial immediately at temperatures below -150 degrees. If you plan to culture the cells immediately, swiftly thaw the vial by shaking it in a 37 degrees water bath with clean water and an antimicrobial agent for 40-60 seconds. Remove the vial once a small ice clump persists, ensuring it remains cold. Proceed with all subsequent steps under aseptic conditions. In a sterile flow hood, disinfect the cryovial with 70% ethanol. Then, gently open the vial and transfer the cell suspension into a 15 ml centrifuge tube pre-filled with 8 ml of room temperature culture medium. Gently mix the cells. For cell separation, centrifuge at 300 x g for 3 minutes and dispose of the supernatant. Skipping centrifugation is optional, although any residual freezing medium should be removed after 24 hours. Resuspend the pellet gently in 10 ml of fresh culture medium and divide between two T25 culture flasks. Follow the subculture protocol for subsequent steps. |
Quality control of H1299 cells
Sterility | Mycoplasma contamination is rigorously 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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