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NCI-H441 [H441]


NCI-H441 [H441] cells were isolated in 1982 from the pericardial fluid of a male patient with papillary adenocarcinoma of the lung. Use these cells as a transfection host for expression of pulmonary surfactant protein (SP-B).
Product category
Human cells
Homo sapiens, human
Adenocarcinoma; Papillary
3D cell culture
High-throughput screening
Product format
Storage conditions
Vapor phase of liquid nitrogen
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ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information


Specific applications
The line has been used as a transfection host for expression of pulmonary surfactant protein (SP-B).


Growth properties
The NCI-H441 cell line was derived by A.F. Gazdar, M. Brower and D. Carney and associates in 1982 from the pericardial fluid of a patient with papillary adenocarcinoma of the lung.
Clinical data
The NCI-H441 cell line was derived by A.F. Gazdar, M. Brower and D. Carney and associates in 1982 from the pericardial fluid of a patient.
modal number = 52; range = 44 to 59.
This is a hyperdiploid human cell line. The modal chromosome number was 52, but cells with 53 chromosome counts also occurred at a high frequency. The rate of cells with higher ploidies was 4.9%. Over 14 marker chromosomes were common to all cells, and 6 or more others were present in some cells. Among the common markers were: der(1)t(1;1)(p36;p21), i(2q), der(11)t(11;12)(q21;q13), t(6p12q), der(9)t(9;?;14)(p24;?q11), t(12q22q) and 8 or more others. All these markers were present in a single copy per cell. Structurally normal N13 and N14 were absent; and N1, N6, N9, N12, N17, F and G chromosomes were single. A single copy each of the X and Y chromosome was detected. The presence of the Y chromosome was confirmed by fluorescent microscopy.
AK-1, 1
ES-D, 1
GLO-I, 2
Me-2, 1
PGM1, 1
PGM3, 1
The cell line expresses mRNA and protein of the major surfactant apoprotein (SP-A).
Electron microscopy shows multilamellar bodies and cytoplasmic structures resembling clara cell granules.
The cells can be cloned in soft agar with or without serum.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, ATCC 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) to a final concentration of 10%.
Handling procedure

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  3. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete growth medium and spin at approximately 125 xg for 5 to7 minutes.
  4. Resuspend cell pellet with the recommended complete growth medium (see the specific batch information for the culture recommended dilution ratio). and dispense into a 25 cm2 or a 75 cm2 culture flask.  It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
  5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.
Subculturing procedure
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
    Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
    Medium Renewal: 2 to 3 times per week
    Note: If desired, the serum can be reduced to 5%.
    Reagents for cryopreservation
    Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)

    Quality control specifications

    Mycoplasma contamination
    Not detected
    Population doubling time
    Approximately 58 hrs in medium with serum; approximately 99 to 138 hrs in serum-free medium
    STR profiling
    D3S1358: 18
    TH01: 9.3
    D21S11: 32.2
    D18S51: 18,19
    Penta_E: 12
    D5S818: 11,12
    D13S317: 9
    D7S820: 10
    D16S539: 9,13
    CSF1PO: 11,12
    Penta_D: 10,12
    Amelogenin: X,Y
    vWA: 17
    D8S1179: 8,14
    TPOX: 8,10
    FGA: 24,25
    D19S433: 14
    D2S1338: 17,19


    Deposited as
    Homo sapiens
    AF Gazdar, JD Minna
    Year of origin
    Special collection
    Human Tumor Cell Bank

    Legal disclaimers

    Intended use
    This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.

    The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.


    This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

    While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

    This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

    Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at

    Permits & Restrictions

    Import Permit for the State of Hawaii

    If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

    This cell line was deposited by Dr. A. Gazdar and is provided for research purposes only. This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement:

    1. Transfers - Biological Materials may not be transferred to third parties for purposes of sale, or producing for sale
    2. Commercial Use - all for-profit and non-profit Recipients must obtain a commercial use license prior to Commercial Use
    Any proposed Commercial Use with these cells must first be negotiated with:

    National Cancer Institute (NCI)
    Email: [email protected]


    Frequently Asked Questions


    Curated Citations

    Bepler G, et al. Expression of p64c-myc and neuroendocrine properties define three subclasses of small cell lung cancer. Oncogene 4: 45-50, 1989. PubMed: 2536917

    Brower M, et al. Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serum-free defined medium. Cancer Res. 46: 798-806, 1986. PubMed: 3940644

    Broers JL, et al. Spontaneous changes in intermediate filament protein expression patterns in lung cancer cell lines. J. Cell Sci. 91: 91-108, 1988. PubMed: 2473086

    O'Reilly MA, et al. Differential effects of glucocorticoid on expression of surfactant proteins in a human lung adenocarcinoma cell line. Biochim. Biophys. Acta 970: 194-204, 1988. PubMed: 3382698

    O'Reilly MA, et al. In vitro translation, post-translational processing and secretion of pulmonary surfactant protein B precursors. Biochim. Biophys. Acta 1011: 140-148, 1989. PubMed: 2713400

    View All Curated Citations for this Product

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