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T cells play a central role in cell-mediated immunity, mediating long-term, antigen-specific, effector, and memory responses. T cells are targets for many immunotherapies, including checkpoint inhibitors, bispecific T-cell engagers, and immune agonists. Furthermore, T cells themselves have been studied as therapeutic agents using chimeric antigen receptor (CAR) addition or autologous adoptive transfer in hematological malignancies. In recent years, various immunotherapeutic strategies aimed at inducing, enhancing, and/or engineering T cell responses have emerged as promising approaches for treating diseases such as cancer and autoimmune disorders.

  
The T cell receptor (TCR) is the molecular structure that enables T cells to specifically recognize and bind antigen peptide-MHC molecules. The activation of T cells mediated by the TCR plays a crucial role in thymocyte T cell development, T cell subset differentiation, and the functional exertion of effector T cells. The TCR can specifically recognize antigen peptides presented on the surface of antigen-presenting cells via MHC molecules, converting extracellular recognition into signals transmissible to the interior of the cell. This process activates tyrosine kinases adjacent to the TCR, promotes the assembly of signal transduction complexes, activates downstream signaling pathways such as MAPK, PKC, and calcium ions, ultimately activating corresponding transcription factors, regulating the expression of effector protein molecules, and completing T cell activation.

The CHO-K1 Human TCR Activator Cell model effectively mimics the in vivo TCR signaling transduction process. The principle is illustrated in the figure below.

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

Figure 2. Response of NFAT-Luc Jurkat (C7C17) to TCR Activitor CHO（C5).

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 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.