The line is available with the following restrictions: 1. This cell line was deposited at the ATCC by Dr. A. Gazdar and Dr. J. Minna 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. 2. Any proposed commercial use of the these cells, or their products must first be negotiated with the University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Blvd., Dallas, Texas 75235. Telephone (214) 699-8056, FAX (214) 688-7233.

transfection host (technology from amaxa
Roche FuGENE® Transfection Reagents)

male

The cells have a homozygous partial deletion of the p53 protein, and lack expression of p53 protein. They reported to be able to synthesize the peptide neuromedin B (NMB) at 0.1 pmol/mg protein, but not the gastrin releasing peptide (GRP).

Protocol:

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 37C 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 37C.
   
   
   Subcultivation ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
   
   Medium renewal: Every 2 to 3 days
   

Recommended medium (without the additional supplements or serum described under ATCC Medium): ATCC 30-2001
recommended serum: ATCC 30-2020
purified DNA: ATCC CRL-5803D