NCI-H1648 [H1648] (ATCC® CRL-5882™) Organism: Homo sapiens, human / Cell Type: lymphoblast / Tissue: lung; derived from metastatic site: lymph node / Disease: stage 3A,adenocarcinoma General Information Characteristics Culture Method Specifications History Documentation Print Email Share Share this product Facebook Twitter Google+ Permits and Restrictions View Permits View Restrictions Organism Homo sapiens, human Tissue lung; derived from metastatic site: lymph node Cell Type lymphoblast Product Format frozen Morphology epithelial Culture Properties adherent 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 3A,adenocarcinoma Age 39 years Gender male Ethnicity Black Derivation The line was established in May 1987. Clinical Data 39 years Black male The patient was a smoker. Comments A lymphoblastoid line from the same patient is available as ATCC CRL-5954. Complete Growth Medium ACL-4 medium (serum-free)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: 0.02 mg/ml insulin 0.01 mg/ml transferrin 25 nM sodium selenite (final conc.) 50 nM Hydrocortisone (final conc.) 1 ng/ml Epidermal Growth Factor (do not filter) 0.01 mM ethanolamine (final conc.) 0.01 mM phosphorylethanolamine (final conc.) 100 pM triiodothyronine (final conc.) 0.5% (w/v) bovine serum albumin (final conc.) 10 mM HEPES 0.5 mM sodium pyruvate (final conc.) extra 2mM L-glutamine (for final conc. of 4.5mM) Subculturing Volumes used in this protocol are for 75 cm2 flasks; 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution . 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 2.0 to 3.0 mL of complete growth medium and aspirate cells by gently pipetting. Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Subcultivation Ratio: Subcultivation Ratio: 1:3 to 1:6 Cryopreservation Culture medium, 95%; DMSO, 5% STR Profile Amelogenin: X,YCSF1PO: 10,12D13S317: 12D16S539: 11D5S818: 11D7S820: 10,11THO1: 7,9.3TPOX: 8,11vWA: 14,17 Name of Depositor AF Gazdar, JD Minna Deposited As Homo sapiens Year of Origin 1987 References NCI-Navy Medical Oncology Branch Cell Line Supplement. J. Cell. Biochem. suppl. 24: 1996. Permits These permits may be required for shipping this product: Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required 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.