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The cytokine Thymic Stromal Lymphopoietin (TSLP) is a type I cytokine that serves as a key mediator of type 2 immune responses and acts as an initiator of T helper 2 (Th2) cell-mediated diseases, including asthma and atopic dermatitis (AD). It plays a critical role in the maturation of T cell populations by activating antigen-presenting cells. The production of TSLP can be induced by a variety of triggers, including tissue injury; ligands for TLR2 (Toll-like receptor 2), TLR3, and NOD2; parasitic, bacterial, and viral infections; proteases (such as trypsin and papain); and pro-inflammatory cytokines.  

TSLP binds to a heterodimeric receptor complex composed of TSLPR (Thymic Stromal Lymphopoietin Receptor) and the IL-7 receptor alpha chain (IL-7Rα), thereby initiating a phosphorylation cascade mediated by JAK1 and JAK2. The subsequent recruitment and activation of STAT5A and STAT5B ultimately lead to the transcription and production of Th2-associated cytokines, specifically IL-4, IL-5, IL-9, and IL-13.  

The TSLP Effector Reporter Cell model accurately recapitulates the in vivo TSLP signal transduction pathway; the underlying principle is illustrated in the figure below.

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

Figure 2. Recombinant hTSLP Effector Reporter Cell stably expressing TSLPR& IL7Rα.

Figure 3. Dose Response of Recombinant Human TSLP in TSLP Effector Reporter Cell(C10).

Figure 4. Inhibition of hTSLP induced Reporter Activiy by hTSLP Neutralizing Ab in TSLP Effector Reporter Cell(C10).

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 TSLP 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.