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HNO258
Tissue
Oral cavity
Disease
Head and neck squamous cell carcinoma (HNSCC)

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HNO258

Cell line · Product number 300146

The HNO258 cell line is derived from an oral squamous cell carcinoma, which is a subtype of head and neck squamous cell carcinoma (HNSCC). This cell line ...

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01General information

Description

The HNO258 cell line is derived from an oral squamous cell carcinoma, which is a subtype of head and neck squamous cell carcinoma (HNSCC). This cell line exhibits several chromosomal abnormalities, which have been identified through chromosomal comparative genomic hybridization (cCGH). Specifically, HNO258 has shown DNA copy number gains in the chromosomal regions 1q41, 3q21-qter, 7p, 7cen-q21, 8q22-qter, 9cen-p13, 9q31-qter, 11q13, 15p, and 15q. Additionally, it displays copy number losses in the regions 4p and 18q12-qter. These genetic alterations are common in HNSCC and are associated with tumorigenesis and cancer progression.

The amplification of 11q13, observed in HNO258, is particularly noteworthy due to its association with the overexpression of oncogenes such as CCND1 (cyclin D1) and CTTN (cortactin), which are involved in cell cycle regulation and cytoskeletal organization, respectively. These oncogenes are frequently implicated in the aggressive behavior of cancer cells, contributing to increased proliferation and invasiveness. The detailed genetic characterization of HNO258 makes it a valuable model for studying the molecular mechanisms underlying oral squamous cell carcinoma and for evaluating potential therapeutic strategies that target these specific genetic alterations.

Tissue
Oral cavity
Disease
Head and neck squamous cell carcinoma (HNSCC)

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
Epithelial-like

04Identifiers

Citation
HNO258 (Cytion catalog number 300146)
NCBI TaxID
9606
Cellosaurus
CVCL_D221

05Handling and culture

Culture medium
DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
Incubation atmosphere

37°C, 5% CO2, humidified atmosphere.

Supplements
Supplement the medium with 10% FBS
Dissociation reagent
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
Thawing and culturing
  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.

06Quality control and cryopreservation

Sterility

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.

Freeze medium
As a cryopreservation medium, we 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.
Storage conditions

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.

Shipping conditions

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.

07Donor

Donor age
62 years
Ethnicity
Caucasian