Isolation date: 1982

Amelogenin: X,Y
CSF1PO: 13,14
D13S317: 8
D16S539: 12
D5S818: 11
D7S820: 10,11
THO1: 8,9.3
TPOX: 9,11
vWA: 18,19

AK-1, 1; ES-D, 1; G6PD, B; GLO-I, 2; Me-2, 0; PGM1, 1-2; PGM3, 1

male

The NCI-H446 cell line was derived by D. Carney, A.F. Gazdar and associates in 1982 from the pleural fluid of a patient with small cell cancer of the lung.
The morphology of the original tumor was not characteristic of SCLC.
The line is a biochemical and morphological variant of SCLC that expresses neuron specific enolase and the brain isoenzyme of creatine kinase.
It does not have detectable levels of L-DOPA decarboxylase, bombesin, vasopressin, oxytocin or gastrin releasing peptide.
C-myc DNA sequences are amplified about 20 fold, and there is a 15 fold increase in c-myc RNA relative to normal cells.
The line was originally propagated in serum free RPMI 1640 medium supplemented with 10 nM hydrocortisone, 0.005 mg/ml insulin, 0.01 mg/ml transferrin, 10 nM 17-beta-estradiol, and 30 nM sodium selenite, 95%.
Fetal bovine serum, 5%.

Protocol: This is a cell line that grows as both attached and suspended cells. The suspended cells are viable and can be used for subculture.To subculture the attached cells, remove the old medium (recover the suspended cells by centrifugation if desired), rinse the monolayer with fresh 0.25% trypsin, 0.53 mM EDTA and let the culture sit at 37C until the cells detach. Add fresh medium, disperse cells and transfer to a new flask.

Subcultivation ratio: A subcultivation ratio of 1:3 to 1:9 is recommended

Medium renewal: 2 to 3 times per week

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