|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.|
|Disease||Lung small cell carcinoma|
|Metastatic site||Pleural Effusion|
|Synonyms||NCI-H446, H-446, NCI-446, NCIH446|
Identifiers / Biosafety / Citation
|Citation||H446 (Cytion catalog number 305049)|
Expression / Mutation
|Tumorigenic||Yes, in nude mice (The cells form transplantable tumors with non-typical SCLC histology).|
|Culture Medium||RPMI 1640|
|Medium supplements||10% FBS, w: 2.1 mM stable Glutamine, w: 2.0 g/L NaHCO3|
|Subculturing||Collect suspension cells in a 15 ml tube and carefully rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add Accutase (1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 10 minutes, then centrifuge the cells growing in suspension and the adherent cells together. Carefully resuspend the cells and dispense into new flasks which contain 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) or CM-ACF (Cytion catalog number 806100)|
|Handling of cryopreserved cultures||The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150 degree Celsius after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 degree Celsius water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the subculture section.|
|Handling of proliferating cultures||One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 37 degree Celsius for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 37 degree Celsius for a minimum of 24 hours.|
Quality control / Genetic profile / HLA
|Sterility||Mycoplasma contaminations are ruled out through PCR-based and luminescence-based mycoplasma assays. The absence of bacterial, fungal or yeast contamination is controlled through daily visual cell monitoring.|
Penta E: 9,10
Penta D: 12,13
To identify mycoplasma contaminations we perform PCR-based and luminescence-based mycoplasma assays. We further determine any bacterial or fungal contamination through our standardized manufacturing processes.
Besides genomic DNA, RNA, cell pellets, and cell lysates, we can offer large quantities of assay-ready cells, plated cells in multiple formats, and frozen or growing cells. Contact us to receive a quote.
Each manufactured batch of cell lines* is authenticated via STR analysis. Contact us if you require a publication-ready STR report for your cells (*human, hamster, mouse, rat, and dog cells).
HLA characterization is available from more than 200 cell lines. HLA class I -A, B, C, and Class II HLA-DPA1, -DPB1, -DQA1, DQB1, and DRB1 alleles were obtained by next-generation sequencing methodologies (NGS) for class I and class II alleles.