||Lung; derived from metastatic site: lymph node.
||Clusters in suspension
||This line was derived by A.F. Gazdar, H.K. Oie, J.D. Minna and associates from a lymph node metastasis obtained from a patient prior to therapy.
||Neuromedin B (NMB)
||The cells are able to synthesize the peptide NMB at 0.2 pmol/mg protein, but not the gastrin releasing peptide (GRP).
||DMEM: F-12 Medium contains 2.5 mM L-glutamine, 15 mM HEPES, 0.5 mM sodium pyruvate, and 1200 mg/L sodium bicarbonate with:
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)
||Cultures can be maintained by addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation of the suspension with subsequent resuspension in fresh medium. Add medium as the cell density increases.
||Atmosphere: air, 95%; carbon dioxide (CO2), 5%
||95% FBS + 5% DMSO (Dimethyl sulfoxide)
|Thawing Frozen Cells:
||SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that
some vials 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 vessel exploding or blowing off its cap with dangerous force creating flying debris.
1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring 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. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. 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 7 minutes.
4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio).
5. Incubate the culture in a appropriate atmosphere and temperature (see ""Culture Conditions"" for this cell line).
NOTE: 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 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).
||Giaccone G, et al. Neuromedin B is present in lung cancer cell lines. Cancer Res. 52: 2732s-2736s, 1992. PubMed: 1563005
NCI-Navy Medical Oncology Branch Cell Line Supplement. J. Cell. Biochem. suppl. 24: 1996.
||Rui Manoel Reis - Hospital de Câncer de Barretos