NCI-H2452

The NCI-H2452 cell line is a human malignant pleural mesothelioma cell line, which was derived from the pleura of a patient with mesothelioma. It is frequently used in research focused on understanding mesothelioma pathophysiology and developing new therapeutic approaches. Like other mesothelioma cell lines, NCI-H2452 is associated with exposure to asbestos fibers, a well-established risk factor for mesothelioma. Studies involving NCI-H2452 have highlighted its utility in exploring mechanisms of disease progression and response to various therapies, particularly gene therapies and viral oncolysis approaches.

NCI-H2452 cells express Coxsackie and adenovirus receptor (CAR) and CD46, which makes them suitable candidates for adenovirus-based gene therapy studies. In research investigating oncolytic virotherapy, both adenovirus type 5 (Ad5) and a fiber-modified variant (Ad5F35) have been tested on NCI-H2452 cells. These adenoviruses replicate selectively within tumor cells, inducing oncolysis in a viral particle-dependent manner. It was found that both Ad5 and Ad5F35 displayed similar efficacy in inducing cell death in NCI-H2452 cells, supporting their potential in gene therapy for malignant mesothelioma.

In addition to its role in oncolytic virotherapy, NCI-H2452 cells have been used to study tumor angiogenesis, a key factor in mesothelioma progression. NCI-H2452 expresses progranulin (PGRN) and granulin-like proteins, which have been identified as novel angiogenic factors that operate independently of the VEGF pathway. This VEGF-independent angiogenesis is crucial, as it offers alternative therapeutic targets in cases where anti-VEGF therapies like bevacizumab fail to improve patient outcomes. Research indicates that these granulins contribute significantly to the formation of new blood vessels, which supports tumor growth and may be involved in the resistance to certain treatments.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. 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.

3. 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.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. 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.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. 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.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.