NCI-H661 Cell Line

43 years

The line lacks ultrastructural and biochemical evidence of squamous differentiation or mucin production.The cells express easily detectable p53 mRNA at levels comparable to normal lung tissue, and exhibit no gross structural DNA abnormalities.The cells stain positively for keratin and vimentin but are negative for neurofilament triplet protein.

BSL-1

Cancer cell line

Freeze medium: 60% Basal medium+30% FBS+10% DMSOStorage temperature: Liquid nitrogen vapor phase

RPMI-1640(PM150110)+10% FBS+1% P/S(PB180120)

~60 hours

Adherent

Atmosphere: Air, 95%; CO2, 5%Temperature: 37°C

37°C

Every 2 to 3 days

Epithelial-like

1:3-1:4

1:2 to 1:4

Remove and discard culture medium. Briefly rinse the cell layer with DPBS solution to remove all traces of serum that contains trypsin inhibitor. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 2 to 3 minutes). Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 4.0 to 6.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels.

Lung