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The TCRα/β-CD3 gene is the core molecular complex for T cells to recognize antigens and initiate immune responses. TCRα/β is a heterodimer composed of α and β chains, belonging to the immunoglobulin superfamily. Its variable region contains three hypervariable regions: CDR1, CDR2, and CDR3. CDR3 exhibits high diversity due to gene rearrangement and N nucleotide insertion, directly determining antigen-binding specificity and recognizing antigen peptide-MHC complexes. The CD3 complex is composed of γ, δ, ε, and ζ subunits, containing 10 ITAM motifs. It tightly associates with TCR through electrostatic interactions in the transmembrane region, forming a TCR-CD3 octamer structure. This complex transmits the antigen signal recognized by TCR into the cell via ITAM phosphorylation, activating kinases such as Lck and ZAP-70, triggering Ca²⁺ release, NF-κB/AP-1/NFAT pathway activation, and effector gene transcription, promoting T cell proliferation and differentiation into effector T cells to carry out cellular immune functions. TCR gene rearrangement (such as V-J, V-D-J recombination) endows an individual's T cell repertoire with diversity and forms the basis of adaptive immunity.

Figure 1. Recombinant TCRα/β-CD3 complex CHO stably expressing TCRα/β-CD3.

Figure 2. Recombinant TCRα/β-CD3 complex CHO stably expressing CD3G&CD3E.

Figure 3. Recombinant TCRα/β-CD3 complex CHO stably expressing CD3D&CD3E.

Figure 4. Recombinant TCRα/β-CD3 complex CHO stably expressing CD3Z.

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 TCRα/β-CD3 complex 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.