NCI-H446 Cells

Name: CLS Cell Lines Service GmbH
Address: Dr.-Eckener-Straße 8, Eppelheim, 69214, DE
Telephone: +496221405780
Price range: €€€

€375.00*

Prices excl. taxes plus shipping costs

Product number: 305049

Safety data sheet

Product sheet

Description	This cell line was established in 1982 by D. Carney, A.F. Gazdar and associates from the pleural fluid of a patient with small cell cancer of the lung. The original tumor morphology was not characteristic of small cell lung cancer. The cell line is a variant small cell lung cancer in biochemistry and morphology, and expresses neuron specific enolase as well as the brain isoenzyme of creatine kinase. None of L-DOPA decarboxylase, bombesin, vasopressin, oxytocin or gastrin releasing peptide has been detected in the cell line. This cell line exhibits a 20-fold higher degree of c-myc DNA amplification and a 15-fold higher degree of c-myc RNA. The cell line was originally propagated in serum free RPMI 1640 medium supplemented with 10 nM of hydrocortisone, 5 microgram/mL of insulin, 10 microgram/mL of transferrin, 10 nM of 17-beta-estradiol, and 30 nM of sodium selenite. Transplantable tumors with non-typical small cell lung cancer histology can be formed by the cells.
Organism	Human
Tissue	Lung
Disease	Lung small cell carcinoma
Metastatic site	Pleural Effusion
Synonyms	NCI-H446, H-446, NCI-446, NCIH446

Age	61 years
Gender	Male
Ethnicity	European
Morphology	Epithelial-Like
Growth properties	Adherent/suspension

Citation	NCI-H446 (Cytion catalog number 305049)
Biosafety level	1

Tumorigenic	Yes, in nude mice (The cells form transplantable tumors with non-typical SCLC histology).

Culture Medium	RPMI 1640, w: 4.5 g/L Glucose, w: 2 mM L-Glutamine, w: 10 mM HEPES, w: 1 mM Sodium pyruvate, w: 1.5 g/L NaHCO3 (Cytion article number 820702a)
Medium supplements	Supplement the medium with 10% FBS
Passaging solution	Accutase
Subculturing	Gather the suspension cells in a 15 ml tube and gently wash the adherent cells with PBS lacking calcium and magnesium (use 3-5 ml for T25 flasks and 5-10 ml for T75 flasks). Apply Accutase (1-2 ml for T25 flasks, 2.5 ml for T75 flasks) ensuring full coverage of the cell layer. Allow the cells to incubate at room temperature for 10 minutes. Following incubation, combine and centrifuge both the suspension and adherent cells. After centrifugation, carefully resuspend the cell pellet and transfer the cell suspension into new flasks containing fresh medium.
Split ratio	1: 3 to 1: 4
Fluid renewal	2 to 3 times per week
Freeze medium	CM-1 (Cytion catalog number 800100)
Handling of cryopreserved cultures	Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Sterility	Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.
STR profile	Amelogenin: x,x CSF1PO: 13 D13S317: 8 D16S539: 12 D5S818: 11 D7S820: 10,11 TH01: 8,9.3 TPOX: 9,11 vWA: 18,19 D3S1358: 17 D21S11: 28 D18S51: 12,13 Penta E: 9,1 Penta D: 12,13 D8S1179: 13,15 FGA: 22 D1S1656: 14,16.3 D6S1043: 11 D2S1338: 18,2 D12S391: 17,18 D19S433: 13,14

NCI-H446 Cells

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