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BAFF (B-cell activating factor), also known as TNFSF13B (tumor necrosis factor superfamily member 13B) or BLyS (B-lymphocyte stimulator), is a cytokine belonging to the tumor necrosis factor (TNF) superfamily. Primarily produced by monocytes, macrophages, dendritic cells, and certain stromal cells, BAFF plays a critical role in B-cell survival, proliferation, and maturation.

BAFF regulates B-cell development and function through receptor binding, and its overexpression is closely associated with autoimmune diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). BAFF has three primary receptors: BAFF-R (BR3), TACI, and BCMA. Binding of BAFF to BAFF-R activates the NF-κB and PI3K/AKT pathways, promoting B-cell survival and proliferation; TACI mediates signaling for both BAFF and APRIL (another TNF family member), regulating B-cell activation and antibody production; and BCMA is primarily expressed on plasma cells, where it mediates signaling for BAFF and APRIL to support plasma cell survival. Overexpression of BAFF is closely linked to autoimmune conditions such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and Sjögren's syndrome.

Serving as target cells for TACI or BAFFR effector reporter cells, BAFF-expressing CHO cells effectively mimic the TACI/BAFF or BAFFR/BAFF signaling transduction processes that occur in vivo, the principle is illustrated in the figure below.

Figure 1. Schematic diagram of TACI & BAFF signal transduction models

Figure 2. Schematic diagram of  BAFFR&BAFF signal transduction models

Figure 3. Recombinant BAFF CHO stably expressing BAFF.

Figure 4. Blocking of BAFF induced TACI Effector Reporter Cell(C32) Activity by BAFF Neutralization Ab with BAFF CHO.

Figure 5. Blocking of BAFF induced BAFFR Effector Reporter Cell(C14) Activity by BAFFR Blocking Ab with BAFF CHO. Blocking of BAFF induced BAFFR Effector Reporter Cell(C14) Activity by BAFFR Blocking Ab with BAFF CHO.

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  CHO-K1 Human BAFF 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.