H9 (derivative of HuT 78)

The H9 cell line, derived from a clonal derivative of the HUT 78 T-cell line from an adult patient with Sezary syndrome, exhibits specific clinical characteristics making it highly relevant in HIV research. It is notably permissive for HIV-1 replication, facilitating the isolation and propagation of HIV-1 from the blood of patients with AIDS and pre-AIDS conditions. This feature underscores its utility in studying viral behaviors and testing antiviral strategies under varied clinical scenarios.

Karyotypically, H9 is near triploid with a modal chromosome number of 69, ranging from 58 to 74, and exhibits a 2.5% frequency of higher ploidies. The cell line displays an extremely complex karyotype, with nearly 60% of the chromosomes per cell consisting of structurally altered marker chromosomes, including translocations such as t(3p4q), t(5q6q), t(5p6p), and deletions like del(7)(q32). Such chromosomal abnormalities contribute to the line’s unique genetic profile, influencing its behavior and response to viral infections. The absence of normal chromosomes N4, N5, N6, N7, N10, N13, N18, N19, N20, and X further distinguishes its genetic makeup.

Moreover, the H9 cell line is tumorigenic in nature, demonstrated by successful subcutaneous tumor formation in nude mice when inoculated with 10(7) cells. It expresses a range of antigens including CD4 and various human leukocyte antigens (HLA) like A1, B62, C3, DR4, and DQ3, which play critical roles in immune recognition and response. Its susceptibility to HIV-1 and expression of genes like interleukin-2 (IL-2) are pivotal for investigating immune responses and viral interactions, making H9 a vital tool in the landscape of immunological and virological research.

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.