HCC366

HCC366 is a human lung adenosquamous carcinoma cell line established from the malignant pleural effusion of an 80-year-old European female patient at the Hamon Cancer Center, UT Southwestern Medical Center. Adenosquamous carcinoma is a rare mixed histological subtype of non-small cell lung cancer (NSCLC) combining features of both adenocarcinoma and squamous cell carcinoma. HCC366 is is included in the Cancer Dependency Map (DepMap), CCLE, COSMIC cell lines, and MD Anderson Cell Lines Project panels. The line carries a homozygous TP53 p.Tyr220Cys mutation (a structural mutation of the DNA-binding domain) and a heterozygous ATM p.Pro534Ala mutation. It does not carry KRAS, EGFR, or STK11 driver mutations, and exhibits microsatellite stability (MSS). The doubling time is approximately 60–70 hours.

HCC366 is applicable in NSCLC research, particularly in studies of lung adenosquamous carcinoma biology, which remains understudied relative to pure adenocarcinoma or squamous cell carcinoma subtypes. Key applications include: TP53 gain-of-function mutation studies; ATM-mediated DNA damage response; chemotherapy sensitivity and resistance testing (cisplatin, paclitaxel, gemcitabine); evaluation of targeted agents; drug sensitivity profiling within DepMap and CCLE frameworks; biomarker discovery for mixed histology NSCLC; comparative genomics; and proteomic/transcriptomic profiling. HCC366 has also contributed to studies of lung cancer cell biology including proliferation, migration, and invasion mechanisms relevant to malignant pleural disease.

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.