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Tumor Necrosis Factor-alpha (TNF-α) is a transmembrane protein with a molecular weight of 26 kDa; upon cleavage of its precursor peptide by TNF-α-converting enzyme (TACE), it is secreted as a soluble 17 kDa molecule. Although activated macrophages are the primary source of TNF-α, it can also be produced by a variety of other cell types—including fibroblasts, astrocytes, smooth muscle cells, keratinocytes, and various tumor cells (such as B-cell lymphomas, breast cancer cells, and colon cancer cells).

Acting as a tumor-promoting factor, TNF-α participates in every stage of tumorigenesis, including cellular transformation, proliferation, angiogenesis, invasion, and metastasis. Furthermore, TNF-α signaling constitutes a critical component of the immune system, capable of suppressing tumorigenesis and inhibiting viral replication; it also functions as an endogenous pyrogen, capable of inducing fever and apoptosis.

Membrane-Anchored TNF-α Cells (Suspension) stably express human TNF-α.

Figure 1. Recombinant Membrane Anchored TNFα Cell (Suspended) constitutively expressing TNFα.

Figure 2. Dose response of TNFα Abs in ADCC Bioassay Effector Cell V variant (High Affinity)-Fcγ-NFAT Jurkat (C36) with Membrane Anchored TNFα Cell (Suspended, C5).

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.