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Herpesvirus Entry Mediator (HVEM), also known as Tumor Necrosis Factor Receptor Superfamily Member 14 (TNFRSF14), is a human cell surface receptor belonging to the TNF receptor (Tumor Necrosis Factor) superfamily. HVEM is expressed in various immune cells, including T cells, B cells, natural killer cells, dendritic cells, and endothelial cells. It also participates in tumor immune evasion through its frequent expression on melanoma cells.

  
HVEM has four distinct ligands—TNFSF14, LTA, BTLA, and CD160—that play a key role in signal transduction. As a molecular switch, HVEM can interact with its LIGHT family members to activate the transcription factor NF-κB, thereby increasing inflammatory responses and enhancing immune responses. Additionally, it is capable of inducing signaling cascades that promote either cell survival or apoptosis through their death domains or TRAF mediators.

HVEM TCR Activator CHO, which constructs HVEM and TCR Activator together on the CHO cells.

Figure 1. Schematic diagram of the CHO-K1 Human HVEM/TCR Activator Cell model.

Figure 2. Recombinant HVEM/TCR Activator/CHO constitutively expressing HVEM.

Figure 3. Dose Response of BTLA/HVEM Blocking Antibody in BTLA Effector Reporter Cells (C1) With HVEM  TCR Activator CHO.

Cell Resuscitation
1)Rapidly thaw the frozen cells in a 37 °C water bath for approximately 60 seconds. Once thawed (which may take slightly less or more than 60 seconds), immediately transfer the cell suspension from the cryovial into a 15 mL centrifuge tube containing 10 mL of pre-warmed  CHO-K1 Human HVEM/TCR Activator Cell complete culture medium.
2)Centrifuge cells at 1000 rpm for 5 min to remove medium, then resuspend cells in 5 mL of pre-warmed complete medium.
3)Transfer the cell suspension into a T25 culture flask and incubate at 37 °C with 5% CO₂.
4)After approximately 24–36 hours, replace the medium or passage the cells to remove non-adherent dead cells.

Subculturing procedure
1)When the cell density reaches the appropriate confluency for passaging, wash the cells with PBS, then add 1 mL trypsin to detach the cells. When more than 80% of the cells detach upon gently tapping the culture flask, add complete culture medium to terminate digestion. Gently pipette to obtain a single-cell suspension, transfer to a 15 mL centrifuge tube, and centrifuge at 1000 rpm for 5 minutes.

2)Discard supernatant after centrifugation. Resuspend cells in fresh medium to a single-cell suspension and transfer to a new culture flask for continued growth.

Cell Freezing
After trypsinization and centrifugation of cells from each T75 flask or 10 cm culture dish, discard the supernatant. Add 2 mL of cryopreservation medium (90% FBS + 10% DMSO), gently resuspend thoroughly, and aliquot into two cryovials. Immediately place the cryovials into a controlled-rate freezing container (e.g., Nalgene 5100-0001), fill with isopropanol to the indicated level, and store at −80 °C. After 24 hours, transfer the cryovials to liquid nitrogen for long-term storage.