NCI-H82 Cells

The NCI-H82 cell line was derived by A.F. Gazdar and associates in 1978 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 or bombesin. The cells produce an abnormally sized p53 mRNA (3.7 kb). C-myc DNA sequences are amplified about 25 fold, and there is a 24 fold increase in c-myc RNA relative to normal cells. The cells are reported to express functional ANP receptors, but treatment with ANP does not alter their growth pattern. The cells stain positively for neurofilaments and vimentin. There is expression of v-fes, v-fms, Ha-ras, Ki-ras, N-ras and c-raf 1 mRNAs.

Aggregates in suspension. The cells grow in very large aggregates, which are the only viable cell population in the culture.

41 years

Caucasian

1

Supplement the medium with 10% FBS

2 to 3 times per week

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.

When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.

We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success.

A ratio of 1:2 to 1:5 is recommended

p53 positive

Insulin-like growth factor II receptor (IGF II), atrial natriuretic peptide (ANP)