| Field | Specification |
|---|---|
| Mfr No | |
| Species |
Follicular dendritic cell (FDC)-like cell lines (HK cells) from human tonsils were established to investigate the role of FDC in germinal centers of lymphoid follicles. Initially, HK cells expressed markers like CD21, CD23, DRC-1, CD40, VCAM-1, ICAM-1, and HJ2, but lost DRC-1, CD21, and CD23 within three days of culture. Morphologically and functionally, HK cells are distinct from fibroblasts and have unique growth requirements. They bind to B cells, supporting their proliferation, but not to T cells. Activated T cells, stimulated with anti-CD3 antibodies, bind to HK cells, inducing phenotypic changes and promoting their growth. HK cells preferentially bind and stimulate germinal center (GC) B cells, rescuing them from apoptosis. They enhance B cell proliferation in the presence of anti-mu or anti-CD40. These cells also produce soluble factors that contribute to their costimulatory activity. Phenotypic and functional analyses suggest that HK cells may be derived from FDCs, highlighting their potential role in supporting GC B cell maturation and differentiation. Immortalized versions of these HK/FDC-like cells are now also available, offering a more stable and scalable tool for long-term studies of FDC function and B cell interactions.
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Surface antigens: CD14+, CD40+, ICAM-1+, VCAM-1+
- cultureMedium: EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a)
- supplements: Supplement the medium with 10% FBS and 1% NEAA
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- fluidRenewal: 1 to 2 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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