In Vitro Technologies

Permits and Restrictions	View Permits View Restrictions
Organism	Homo sapiens, human
Tissue	lung; derived from metastatic site: lymph node
Product Format	frozen
Morphology	epithelial
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease	stage 4, adenocarcinoma
Age	68 years
Gender	female
Ethnicity	Caucasian
Storage Conditions	liquid nitrogen vapor phase

Derivation	The line was established in August 1988.
Clinical Data	The patient received prior chemotherapy and radiation therapy. The patient was a smoker. Thirty pack years
Comments	A lymphoblastoid line from the same patient is available as ATCC CRL-5961.

Complete Growth Medium	HITES medium supplemented with 5% fetal bovine serum The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No.30-2006. To make the complete growth medium,add the following components to the base medium 0.005 mg/ml Insulin 0.01 mg/ml Transferrin 30nM Sodium selenite (final conc.) 10 nM Hydrocortisone (final conc.) 10 nM beta-estradiol (final conc.) extra 2mM L-glutamine (for final conc. of 4.5 mM) 5% fetal bovine serum (final conc.)
Subculturing	Volumes used in this protocol are for 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. 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. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Subcultivation Ratio: 1:3 to 1:6Medium Renewal: Two to three times weekly Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.
Cryopreservation	85% Complete Growth Medium + 10% FBS + 5% DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Complete Growth Medium

HITES medium supplemented with 5% fetal bovine serum

0.005 mg/ml Insulin
0.01 mg/ml Transferrin
30nM Sodium selenite (final conc.)
10 nM Hydrocortisone (final conc.)
10 nM beta-estradiol (final conc.)
extra 2mM L-glutamine (for final conc. of 4.5 mM)
5% fetal bovine serum (final conc.)

Subculturing

Volumes used in this protocol are for 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
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.
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37°C.

Subcultivation Ratio: 1:3 to 1:6Medium Renewal: Two to three times weekly

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation

85% Complete Growth Medium + 10% FBS + 5% DMSO.

Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

STR Profile	Amelogenin: X CSF1PO: 10,12 D13S317: 12 D16S539: 12 D5S818: 13 D7S820: 9,11 THO1: 9.3 TPOX: 8 vWA: 14

Name of Depositor	AF Gazdar, JD Minna
Year of Origin	1988
References	NCI-Navy Medical Oncology Branch Cell Line Supplement. J. Cell. Biochem. suppl. 24: 1996. Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC. Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988. Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC. Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.

Name of Depositor

AF Gazdar, JD Minna

Year of Origin

1988

References

NCI-Navy Medical Oncology Branch Cell Line Supplement. J. Cell. Biochem. suppl. 24: 1996.

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.

Permits	These permits may be required for shipping this product to Australia: DAFF Import Permit formerly known as AQIS Import Permit must be obtained and a copy of the permit must be sent to ATCC in advance of shipment.
Basic Documentation	Product Sheet Certificate of Analysis SDS
Restrictions	The line is available with the following restrictions: This cell line was deposited at the ATCC by Dr. Adi F. Gazdar and is provided for research purposes only. Neither the cell line nor products derived from it may be sold or used for commercial purposes. Nor can the cells be distributed to third parties for purposes of sale, or producing for sale, cells or their products. The cells are provided as service to the research community. They are provided without warranty of merchantability or fitness for a particular purpose or any other warranty, expressed or implied. Any proposed commercial use of the these cells, or their products, must first be negotiated with the National Cancer Institute (NCI). For further information, please contact NCI’s Technology Transfer Center at NCI_TTC_Contact@mail.nih.gov or by phone at (240)-276-5514.
References	NCI-Navy Medical Oncology Branch Cell Line Supplement. J. Cell. Biochem. suppl. 24: 1996. Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC. Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988. Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC. Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.

E: care@invitro.com.au
P: 1300 552 003

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NCI-H2009; Lung Adenocarcinoma; Human (Homo sapiens) Item ID: ATCCRL5911

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