Committed to providing comprehensive support for global drug development

Thyroid hormone receptors (TRs) belong to the nuclear receptor superfamily and serve as key molecules mediating the biological functions of thyroid hormones (T3/T4). By regulating the transcription of target genes, they participate in metabolism, growth and development, and tissue homeostasis. TRs exist in two subtypes—TRα and TRβ—and the proteins encoded by these genes contain a DNA-binding domain (DBD) and a ligand-binding domain (LBD). 

Thyroid Hormone Receptor β (THRB, or TRβ) is a member of the nuclear receptor superfamily and acts as one of the primary receptors mediating the effects of thyroid hormones (T3/T4). It plays a pivotal role in energy metabolism, growth and development, and tissue homeostasis, regulating target gene transcription either as a homodimer or as a heterodimer (in conjunction with the Retinoid X Receptor, RXR). Compared to THRA (Thyroid Hormone Receptor α), THRB is highly expressed in the liver, heart, and skeletal muscle, where it predominantly governs lipid metabolism regulation (specifically, cholesterol reduction), thermogenesis, and the modulation of cardiac contractility. Under pathological conditions, mutations in THRB can lead to Resistance to Thyroid Hormone (RTH) syndrome, a condition characterized by symptoms of hyperthyroidism accompanied by abnormally elevated levels of Thyroid-Stimulating Hormone (TSH). Furthermore, dysregulation of THRB signaling has been implicated in metabolic syndrome, obesity, and the progression of thyroid cancer. In recent years, selective agonists targeting THRB (such as Eprotirome and Resmetirom) have emerged as a major focus in the treatment of Non-Alcoholic Steatohepatitis (NASH) and dyslipidemia, largely because they allow for the avoidance of the cardiotoxic side effects associated with THRA activation. Several such drug candidates have already advanced to Phase III clinical trials; however, challenges regarding tissue-specific delivery and long-term safety still require further optimization.

The THRB Luciferase Reporter HEK293 cell line consists of HEK293 cells engineered to express a luciferase reporter gene under the transcriptional control of THRB. Thyroid hormones (T3/T4) are potent endocrine signaling molecules that exert their primary effects by binding to TRs, thereby regulating gene expression and cellular function. The underlying principle is illustrated in the figure below.

Figure 1. Schematic Diagram Illustrating the Principle of the THRB Luciferase Reporter HEK293 Cell Line

Figure 2. WB of  hTHRB Luciferase Reporter HEK293.

Figure 3. Dose Response of T3 in THRB Luciferase Reporter HEK293.

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 THRB Luciferase 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.