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The NCI-H441 cell line, also known as H441, established in 1982 from the pleural effusion of a male patient with papillary adenocarcinoma of the lung, is a well-characterized epithelial adenocarcinoma cell line. These cells are extensively utilized in biological research for their relevance to lung epithelial biology, making them a critical in vitro model for studies on transepithelial transport and the epithelial barrier function.
The NCI-H441 cell line serves as a vital tool in advancing our understanding of pulmonary drug disposition and tumor kinetics. Its use in 3D cell culture models allows for the detailed study of how drugs are absorbed, distributed, metabolized, and excreted within the lung environment, closely mimicking in vivo conditions.
Given their origin and characteristics, NCI-H441 cells are particularly valuable in research focused on the distal lung and related diseases, including lung adenocarcinoma, serving as a stable and relevant cell model for understanding the mechanisms of lung diseases and evaluating potential therapeutic interventions.
NCI-H441 cells are instrumental in 3D cell culture, high-throughput screening, and toxicology studies, providing valuable data on cellular responses and the efficacy of therapeutic agents. A notable application of the human cell line H441 involves their use as a transfection host for expressing pulmonary surfactant protein (SP-B), utilizing the firefly luciferase reporter system, which emphasizes their role in inhalation biopharmaceutics and transepithelial transport studies. This capability, alongside their expression of mRNA and protein for major surfactant apoprotein (SP-A), highlights the cell line's significance in investigating lung function and disorders, especially those affecting surfactant regulation and synthesis.
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Karyotype: The NCI-H441 cell line exhibits a hyperdiploid karyotype, with a modal chromosome number of 52, though variations from 44 to 59 chromosomes have been documented.
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- doublingTime: 58 hours
- 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.
- fluidRenewal: 2 to 3 times per week
- freezeMedium: As a cryopreservation medium, 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.
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