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TNFR1 and TNFR2 are two receptors for tumor necrosis factor-α (TNF-α), jointly mediating various biological functions of TNF. TNFR1 (p55) is widely expressed in almost all cells and primarily mediates inflammatory responses and apoptosis through its intracellular death domain. TNFR2 (p75) expression is relatively limited, mainly on immune cells, and mediates cell survival, proliferation, and immune regulation by activating the NF-κB and AKT pathways.

Figure 1. Recombinant TNFR1&TNFR2 Dual Effector Reporter Cell constitutively expressing TNFR1&TNFR2.

Figure 2. Blocking of TNFα induced TNFR1&TNFR2 Dual Effector Reporter Cell Activity by T'NFα Neutralization Ab or TNFR1/R2 Blocking Abs with Membrane Anchored TNFα Cell (Adherent, C12).

Figure 3. Inhibition of Human Recombinant TNFα Induced Reporter Activity by TNFα Neutralization Ab or TNFR1/R2 Blocking Abs in TNFR1&TNFR2 Dual Effector Reporter Cell.

Cell Passage Procedures

1.This cell line grows in suspension.
2.Upon receipt, cells should be thawed immediately or stored in liquid nitrogen until use.
3.Before thawing, pre-warm the water bath and culture medium to 37 °C, and prepare a small amount of dry ice.
4.Remove the cryovial from storage and transport it to the cell culture laboratory on dry ice.
5.Rapidly thaw the cells in a 37 °C water bath. Once the cells are completely thawed, spray the cryovial with 70% ethanol for disinfection and transfer it to a biosafety cabinet.
6.Add 10 mL of pre-warmed culture medium into a 15 mL centrifuge tube. Transfer the contents of the cryovial into the tube and centrifuge at 1000 rpm for 5 minutes.
7.Carefully discard the supernatant. Resuspend the cell pellet in 5 mL of pre-warmed culture medium by gentle pipetting. Immediately perform cell counting and adjust the cell density to 3–6 × 10⁵ cells/mL based on the counting results, then transfer the cells into a culture flask.
8.Count the cells every 1–2 days. When the cell density exceeds 1 × 10⁶ cells/mL, passage the cells promptly or add fresh culture medium. Maintain the cell density between 2 × 10⁵ and 1 × 10⁶ cells/mL.

Suspension Cell Cryopreservation Procedure:

1.Collect 8 × 10⁶ cells, centrifuge, and discard the supernatant.
2.Add 1 mL of cell freezing medium (90% FBS + 10% DMSO) and gently pipette to mix thoroughly. Transfer the suspension into a cryovial.
3.Immediately place the cryovial into a controlled-rate freezing container (Nalgene 5100-0001), fill with isopropanol up to the indicated level, and store at −80 °C.
4.After 24 hours, transfer the cryovial to liquid nitrogen for long-term storage.