acetylcholine, muscarinic, expressed [23078]

myc +; erb B2 +

near diploid; DM were present in 64% of cells examined

NCI-N87 cells express the surface glycoproteins carcinoembryonic antigen (CEA) and TAG 72, and are L-dopa decarboxylase (DDC) negative. They were minimally positive for vasoactive intestinal peptide (VIP) receptors and lacked gastrin receptors. They were fiound to express receptors for muscarinic cholinergic agents. No evidence of amplification or rearrangements was noted with the N-myc, L-myc, myb and EGF receptor genes. The cell line expressed levels of c-myc and c-erb-B 2 RNA that were comparable to other cell lines.There was no expression of the following genes: N-myc, L-myc, c-cis, IGF-2, or gastrin releasing peptide. NCI-N87 cells have a reported plating efficiency of 4.3%.

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 37°C 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 37°C.

Subcultivation ratio: A subcultivation ratio of 1:3 to 1:4 is recommended

Medium renewal: Two to three times weekly

47 hrs

23078: Park JG , et al. Characteristics of cell lines established from human gastric carcinoma. Cancer Res. 50: 2773-2780, 1990. PubMed:
23570: NCI-Navy Medical Oncology Branch Cell Line Supplement. J. Cell. Biochem. suppl. 24: 1996.