AKATA

The AKATA cell line, derived from Burkitt’s lymphoma, is a widely used model to study Epstein-Barr virus (EBV) latency and reactivation. EBV is a ubiquitous herpesvirus linked to a range of cancers, including Burkitt lymphoma, and typically establishes a latent infection within B cells. In AKATA cells, EBV is maintained in an episomal state with a type I latency program, expressing a limited set of viral genes such as EBNA-1, EBER RNAs, and BamHI-A rightward transcripts (BARTs). This restricted gene expression allows the virus to persist in the host without initiating a full lytic cycle. However, AKATA cells can be triggered to enter the lytic phase, where the virus actively replicates and produces progeny. This reactivation is commonly induced through cross-linking surface immunoglobulins, which makes AKATA cells an excellent tool for studying EBV reactivation dynamics and viral gene regulation.

Research utilizing the AKATA cell line has also examined the impact of chemotherapeutic agents on EBV reactivation. For instance, drugs like etoposide and doxorubicin have been shown to influence viral latency. Etoposide induces apoptosis in AKATA cells but reactivates EBV less effectively than doxorubicin, which promotes higher levels of lytic gene expression and viral progeny production. Additionally, studies involving gene editing techniques, such as CRISPR/Cas9, have explored the role of epigenetic regulators in AKATA cells. For example, knockout of the histone methyltransferase EZH2 in AKATA cells disrupts the maintenance of latency by reducing the trimethylation of histone H3K27, leading to increased expression of both latent and lytic EBV genes, as well as enhanced viral replication and cell proliferation.

AKATA cells also display distinct phenotypic characteristics based on EBV presence, such as increased sensitivity to apoptosis-inducing agents and variations in gene expression related to apoptotic pathways. These differences make EBV-positive AKATA cells a powerful model for dissecting EBV's influence on host cell survival, gene expression, and the virus's lifecycle, particularly in the context of cancer development and potential therapeutic interventions targeting EBV-associated malignancies.

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.