Calu-3 Cell Line

Brief Introduction
Product List

General Information
Organism	Homo sapiens, human
Cell Line Description	Calu-3 is a human lung adenocarcinoma cell line established by J. Fogh and colleagues in 1975. It was isolated from primary tissue in the bronchial submucosa of a 25-year-old Caucasian male diagnosed with lung adenocarcinoma. Calu-3 cells exhibit a highly polarized epithelial morphology and grow as an adherent monolayer. In the fields of respiratory biology, toxicology, and infectious disease research, Calu-3 is widely recognized as the preferred in vitro model for studying human upper respiratory tract epithelium. When cultured on permeable Transwell inserts at the air-liquid interface (ALI), Calu-3 cells effectively differentiate into a polarized monolayer, displaying features such as functional tight junctions, microvilli, and active mucin secretion (i.e., a mucociliary-like phenotype). Due to the high, endogenous expression of ACE2 and TMPRSS2 receptors, this cell line has become a vital physiological host model for studying respiratory viruses, including SARS-CoV-2 and influenza strains.
Tissue	Lung
Disease	Lung Adenocarcinoma / Non-Small Cell Lung Cancer (NSCLC)
Morphology	Epithelial-like; polygonal cells growing in tightly packed cohesive clusters
Gender	Male
Age	25 years
Product Format	Frozen
Growth Mode	Adherent
Biosafety Level	1 (Biosafety classification matches standard laboratory parameters for non-infectious human tumor cells)
Applications	1. Highly cited host models for studying the replication kinetics and entry mechanisms of SARS-CoV-2, MERS-CoV, and influenza viruses; 2. Key in vitro models for pulmonary drug delivery, aerosol deposition assays, and the analysis of transepithelial transport characteristics; 3. Use of air-liquid interface (ALI) systems to elucidate human respiratory epithelial barrier function and tight junction assembly mechanisms; 4. Investigation of mucin biosynthesis (specifically MUC5AC and MUC5B expression) and airway inflammatory pathways; 5. Preclinical screening of inhaled therapeutic agents, toxic particulates, and targeted anti-adenocarcinoma drugs.
Shipped In	Dry ice
Storage Temperature	−196°C (Liquid nitrogen vapor phase only)
Characteristics
Tumorigenic	Yes, reliably forms adenocarcinomas when engrafted into standard immunocompromised rodent hosts (e.g., nude mice).
Karyotype	Aneuploid/hyperdiploid; a highly rearranged human karyotype with a modal chromosome number ranging from 72 to 78. It exhibits complex structural marker chromosomes commonly found in advanced-stage pulmonary malignancies.
Genetic Profile	1. TP53 status: Mutant; harboring a hemizygous or homozygous missense mutation at the p.Met237Ile (c.711G>T) site in exon 7. 2. KRAS status: Wild-type. 3. EGFR status: Exhibits high native expression and structural integrity. 4. ERBB2 (HER2) status: Typically characterized by endogenous amplification and high levels of protein expression.
Physiological Markers	Natively synthesizes and secretes high levels of airway mucins, primarily MUC5AC. Expresses polarized junctional proteins including ZO-1 and E-cadherin, generating high transepithelial electrical resistance (TEER) measurements when properly differentiated.
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Remove and discard all used complete culture medium from the culture dish. 2. Briefly rinse the confluent cell monolayer with sterile, Ca2+/Mg2+-free PBS to neutralize residual serum. 3. Add 2.0 to 3.0 mL of 0.25% trypsin-0.53 mM EDTA solution, ensuring the cell monolayer is completely covered. 4. Incubate at 37°C for 5 to 15 minutes, monitoring closely under an inverted microscope. Note: Calu-3 cells are highly adherent, exhibiting strong adhesion to each other and the plastic surface; a full 15-minute incubation may be required to achieve complete detachment into a single-cell suspension. 5. Add an equal volume of pre-warmed complete culture medium to the dish and gently pipette up and down to disperse the cells evenly and break up any clumps. 6. Centrifuge at approximately 125 × g for 5 minutes, discard the supernatant, gently resuspend the cell pellet in fresh complete culture medium, and distribute into new vessels.
Medium Renewal	2 to 3 times per week
Subcultivation Ratio	Split optimal, confluent cultures at a standard ratio of 1:2 to 1:4. Avoid over-diluting the cells, as they depend on paracrine clustering factors to thrive.
Culture Conditions	Atmosphere: Air, 95%; CO2, 5%; Temperature: 37°C
Cryopreservation	90% Complete culture medium + 10% DMSO (or 50% Basal medium + 40% FBS + 10% DMSO)

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

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