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HCC2935

CRL-2869

Product category
Human cells
Organism
Homo sapiens, human
Cell type
epithelial cell
Morphology
epithelial
Tissue
Lung; Pleural effusion
Disease
Adenocarcinoma
Applications
3D cell culture
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Price: $495.00 EA
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Documentation

Biosafety Icon BSL 1

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.

RPMI-1640 Medium

30-2001

Price: $22.00 ea

Fetal Bovine Serum (FBS)

30-2020

Price: $645.00 ea

Trypsin-EDTA Solution, 1X

30-2101

Price: $14.00 ea

Dulbecco's Phosphate Buffered Saline (D-PBS), 1X

30-2200

Price: $16.00 ea

Dimethylsulfoxide (DMSO)

4-X

Price: $49.00 ea

Detailed product information

Characteristics

Growth properties
Adherent
Derivation

Established from pleural effusion cells of an adenocarcinoma patient who had never smoked.

Age
39 years
Ethnicity
White
Gender
Male
Comments
This lung adenocarcinoma has an acquired mutation in the EGFR tyrosine kinase domain (E746 - T751 deletion, S752I).

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%.
Temperature
37°C
Atmosphere
95% Air, 5% CO2
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 culture medium and spin at approximately 125 x g for 5 to 10 minutes.
  4. Resuspend cell pellet with the recommended complete 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.
Subculturing procedure
Volumes used in this protocol are for 75cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution or Dulbecco’s Phosphate Buffered Saline (D-PBS) to remove all traces of serum that contains trypsin inhibitor.
  3. Add 1.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until the 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. Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
  6. Discard supernatant and resuspend cells in fresh growth medium.
  7. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 5 x 103 to 7 x 103 viable cells/cm2 is recommended.
  8. Place culture vessels in incubators at 37°C. Maintain cultures at a cell concentration between 8 x 104 and 1 x 105 cells/cm2

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days

Quality control specifications

Mycoplasma contamination
Not detected
Population doubling time
Approximately 105 hrs
STR profiling
Amelogenin: X,Y
CSF1PO: 12
D13S317: 12
D16S539: 12
D5S818: 13
D7S820: 12,13
THO1: 7,9
TPOX: 8
vWA: 17

History

Deposited as
Homo sapiens
Depositors
JD Minna, AF Gazdar
Year of origin
1999

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.
Warranty

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.

Disclaimers

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 www.atcc.org.

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 Dr. J. Minna 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:

The University of Texas Southwestern Medical Center
5323 Harry Hines Blvd.
Dallas, Texas 75235
Telephone: (214) 648-1888
Email: [email protected]

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Frequently Asked Questions

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Telephone

Telephone

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Outside the US

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