HTB-171 NCI-H446 人小細(xì)胞肺癌細(xì)胞
ATCC® Number: HTB-171?
Designations: NCI-H446 [H446]
Depositors: AF Gazdar, JD Minna
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: mixed, adherent and suspension
Organism: Homo sapiens (human)
Morphology: epithelial
Source: Organ: lung
Disease: carcinoma; small cell lung cancer
Derived from metastatic site: pleural effusion
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Isolation: Isolation date: 1982
Tumorigenic: Yes
DNA Profile (STR): 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
Cytogenetic Analysis: This is a hypertriploid human cell line. The modal chromosome number is 74, occurring at 22% with polyploidy at 2.5%. Over 25 markers were common to most cells including inv(1)(p32q21), der(1)t(1:4)(q42;q11), i(5q), t(2p13q) and der(19)t(7;HSR;19) (q11;HSR;p13.3). Structurally normal N1 and N13 were absent and normal N4, N5 and N10 were mostly single copies per cell. There were two X/Y pairs per cell.
Isoenzymes: AK-1, 1
ES-D, 1
G6PD, B
GLO-I, 2
Me-2, 0
PGM1, 1-2
PGM3, 1
Age: 61 years
Gender: male
Ethnicity: Caucasian
Comments: 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%.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing: 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
Preservation: Freeze medium: Complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001
