CAL-33 Cell Line

Brief Introduction
Product List

General Information
Organism	Homo sapiens, human
Cell Line Description	CAL-33 is a human head and neck squamous cell carcinoma cell line established in 1983 by J. Gioanni and colleagues at the Antoine Lacassagne Center. The cell line was derived from a primary tumor surgically resected from a 69-year-old Caucasian male diagnosed with moderately differentiated squamous cell carcinoma of the tongue prior to receiving chemotherapy or radiotherapy. CAL-33 cells exhibit a characteristic polygonal, epithelial-like morphology and grow as a tightly adherent monolayer. In the fields of oncology, pharmacology, and radiobiology, CAL-33 is a globally recognized, preferred model for HPV-negative head and neck squamous cell carcinoma (HNSCC). It is widely used to elucidate the molecular signaling pathways driving oral malignancies, evaluate novel small-molecule chemotherapeutic agents, and investigate genetic markers of radioresistance.
Tissue	Head and Neck; derived from Oral Cavity (Tongue)
Cell Type	Epithelial cell
Disease	Tongue Squamous Cell Carcinoma (HNSCC)
Morphology	Epithelioid / Polygonal; grows as a flat adherent monolayer
Gender	Male
Age	69 years
Product Format	Frozen
Growth Mode	Adherent
Biosafety Level	1 (Biosafety classification aligns with standard international repository guidelines for non-infectious human tumor cell lines)
Applications	1. Direct in vitro modeling of tumor biology in HPV-negative head and neck squamous cell carcinoma (HNSCC); 2. Standardized models in radiation oncology for evaluating mechanisms of radioresistance, DNA double-strand break repair, and cellular senescence; 3. High-throughput preclinical screening of novel chemotherapeutic agents, targeted kinase inhibitors (e.g., EGFR and PI3K/mTOR inhibitors), and immunotherapies; 4. Investigation of mechanisms underlying tumor cell migration, invasion, and extracellular matrix remodeling (mediated by matrix metalloproteinase secretion); 5. Establishment of robust subcutaneous and orthotopic xenograft models of tongue cancer in immunodeficient mice (e.g., nude or SCID mice)
Shipped In	Dry ice
Storage Temperature	−196°C (Liquid nitrogen vapor phase only)
Characteristics
Tumorigenic	Yes; highly tumorigenic in athymic nude mice, forming progressively expanding squamous cell carcinomas.
Karyotype	Human hyperdiploid karyotype with approximately 4% polyploidy. Modal chromosome distribution center is 49 (ranges from 45–49).
Viral Profile	Verified negative for EBV, HBV, HCV, HIV-1, HIV-2, HPV (Human Papillomavirus), HTLV-1/2, MLV, and SMRV via PCR-based clearance panels.
Microsatellite Status	Stable (MSS).
Growth Kinetics	Robust adherent expansion; typical population doubling time spans approximately 30 to 43 hours depending on culture passage and density.
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Completely remove and discard the used complete culture medium from the culture dish. 2. Gently rinse the cell monolayer with sterile, Ca2+/Mg2+-free PBS to neutralize any residual traces of serum containing active trypsin inhibitors. 3. Add 2.0 to 3.0 mL of pre-warmed 0.25% trypsin-0.53 mM EDTA solution, ensuring the cell layer is completely covered. 4. Incubate at 37°C for 5 to 10 minutes, monitoring closely under an inverted microscope. Note: CAL-33 cells form a highly adherent, tightly packed monolayer; a full 10-minute incubation may be required for the cells to fully round up and detach. 5. Add an equal volume of pre-warmed complete culture medium to rapidly neutralize enzyme activity. 6. Gently pipette the suspension up and down to dissociate the cells, centrifuge at ~125 × g (or 300 × g) for 5 minutes, carefully aspirate the supernatant, gently resuspend the cell pellet in fresh medium, and distribute into new vessels.
Medium Renewal	Every 2 to 3 days (or 2 times per week depending on metabolic density shifts)
Subcultivation Ratio	Split confluent cultures at a standard ratio of 1:3 to 1:5 every 4 to 5 days.
Culture Conditions	Atmosphere: Air, 90%; CO2, 10% (or standard Air, 95%; CO2, 5%); Temperature: 37°C
Cryopreservation	90% Complete culture growth medium + 10% DMSO (or 70% Complete medium + 20% FBS + 10% DMSO)

Cat.No.	Product Name	Price
CSC-RO02608	Cas9 Stable Cell Line - CAL-33	Inquiry
CSC-RR00901	GFP Reporter Cell Line - CAL-33	Inquiry
CSC-RR00949	Luciferase Reporter Cell Line - CAL-33	Inquiry

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* For research use only. Not intended for any clinical use.

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