NCI-H1299

NCI-H1299, also known as H1299, is a human non-small cell lung cancer (NSCLC) cell line established from a lymph node metastasis derived from an adult male patient with lung carcinoma. Along with H292 cells, H1299 is widely used as an NSCLC model in cancer biology and immuno-oncology research. The cell line exhibits an epithelial-like morphology characterized by adherent, flattened cells with a thickness of less than 5 µm and an approximate doubling time of 22–30 hours. H1299 cells express keratin and vimentin but are negative for neurofilament triplet protein, reflecting a phenotype with both epithelial and mesenchymal characteristics.

Genetically, H1299 cells harbor a homozygous partial deletion in the TP53 gene, resulting in complete loss of p53 protein expression. The line is also characterized by wild-type KRAS status, distinguishing it from other NSCLC models such as A549 cells, which carry endogenous KRAS mutations. Due to the absence of functional p53 signaling combined with intact KRAS, H1299 cells are frequently used to study tumor suppressor biology, oncogenic signaling pathways, apoptosis, metastasis, and therapeutic resistance mechanisms. Compared with more epithelial NSCLC cell lines such as A549, H1299 cells display a more mesenchymal phenotype with reduced epithelial marker expression, making them particularly useful for investigations into epithelial-to-mesenchymal transition (EMT), invasion, and metastatic progression.

H1299 cells have also been reported to synthesize the neuropeptide neuromedin B (NMB) at low levels, while lacking detectable gastrin-releasing peptide (GRP) production. Their robust growth characteristics, high transfectability, and well-characterized molecular background have contributed to their broad use in studies involving targeted therapies, gene editing, immune-mediated cytotoxicity, and downstream KRAS-associated signaling pathways. As with all long-term cultured tumor cell models, periodic authentication and confirmation of key molecular features are recommended to ensure experimental reproducibility.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.