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modal number = 46; range = 42 to 48.
This is a pseudodiploid human cell line. The modal chromosome number is 46 occurring at 68% and polyploidy at 0.4%. Three marker chromosomes were detected in all metaphases examined. Two markers that appeared to be the product of the balanced translocation between N1 and N17. The third marker is ?inv(13). No other common markers were detected. Normal N1, N13, and N17 had single-copy each. Y chromosome is present.
The cells express receptors for bombesin at up to 6000 sites per cell.
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Temperature: 37.0°C
Subculturing:
Protocol:
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM 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: A subcultivation ratio of 1:2 to 1:5 is recommended Medium Renewal: 2 to 3 times per week
