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TRKB (Tropomyosin Receptor Kinase B, also known as the BDNF receptor or NTRK2) is a pivotal member of the neurotrophin receptor family (Trk receptors). As a transmembrane tyrosine kinase receptor, it specifically binds to its ligands—Brain-Derived Neurotrophic Factor (BDNF) and Neurotrophin-4 (NT-4). By activating downstream PI3K/Akt, Ras/MAPK, and PLCγ signaling pathways, TRKB serves as a central regulator of neuronal survival, synaptic plasticity, neurotransmitter release, and axon guidance; it acts as a core hub for maintaining learning and memory, emotional balance, and neurodevelopment. In pathological states, TRKB signaling dysfunction is directly implicated in neurological disorders such as major depressive disorder, Alzheimer's disease, neuropathic pain, and epilepsy. Furthermore, as a pan-cancer target, TRKB gene fusion events can drive the progression of solid tumors—including sarcomas and thyroid cancers—for which TRK inhibitors (such as Larotrectinib and Entrectinib) have already been approved as treatments. In the metabolic realm, TRKB is involved in obesity (where hypothalamic TRKB regulates energy balance) and non-alcoholic fatty liver disease (where TRKB in hepatic stellate cells promotes fibrosis). Current targeted strategies against TRKB encompass small-molecule kinase inhibitors (for anti-cancer therapy), BDNF-mimetic peptides (for neuroprotection), and gene therapies (for BDNF delivery); however, these approaches face challenges such as penetrating the blood-brain barrier, achieving tissue selectivity, and overcoming drug-resistance mutations (e.g., TRKB G667C).

The TRKB Effector Reporter Cell model accurately simulates the in vivo signal transduction processes of TRKB; the underlying principle is illustrated in the figure below.

Figure 1. Schematic Diagram of the HEK293 Human TRKB Effector Reporter Cell Model

Figure 2. Recombinant TRKB Effector Reporter Cell stably expressing TRKB.

Figure 3. Dose Response of Recombinant Human NT-4 in TRKB Effector Reporter Cell(C15).Dose Response of TRKB Agonist Ab in TRKB Effector Reporter Cell(C15).

Figure 4. Inhibition of Human NT-4 induced Reporter Activity by NTRK Inhibitor in TRKB Effector Reporter Cell( C15).

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  HEK293 Human TRKB Effector Reporter 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.