KC-1594

CHOK1-LILRB5-Cell-Line

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Background of CHOK1-LILRB5-Cell-Line

Leukocyte immunoglobulin-like receptor subfamily B member 5 (LILRB5), also known as CD85c and LIR-8, belongs to the leukocyte immunoglobulin-like receptor (LIR) family of transmembrane glycoproteins that negatively regulate immune cell activation. LILRB5 transcripts were detected in natural killer cells, B lymphoblasts, mast cells, monocytes, macrophages and osteoclasts, but only mast cells expressed LILRB5 protein. In addition, LILRB5 was also expressed on the surface of peripheral monocytes. LILRB5 specifically binds hLA-B7 and HLA-B27 heavy chains and may play a role in mast cell inflammation. It is expressed in immune cells of patients with liver cancer. No relevant target antibodies have been reported.

Specifications

Catalog Number	KC-1594
Cell Line Name	CHOK1-LILRB5-Cell-Line
Host Cell Line	Chinese hamster ovary CHO-K1 cell line
Description	Stable CHOK1 cell line expressing exogenous human LILRB5 gene
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+10µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1 human LILRB5 cell line was generated using a lentiviral vector expressing the human LILRB5 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 10µg/mL Puromycin)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Zhang Z, Hatano H, Shaw J, Olde Nordkamp M, Jiang G, Li D, Kollnberger S. The Leukocyte Immunoglobulin-Like Receptor Family Member LILRB5 Binds to HLA-Class I Heavy Chains. PLoS One. 2015 Jun 22;10(6):e0129063.
Fan J, Li J, Han J, Zhang Y, Gu A, Song F, Duan J, Yin D, Wang L, Yi Y. Expression of leukocyte immunoglobulin-like receptor subfamily B expression on immune cells in hepatocellular carcinoma. Mol Immunol. 2021 Aug;136:82- 97.
Hogan LE, Jones DC, Allen RL. Expression of the innate immune receptor LILRB5 on monocytes is associated with mycobacteria exposure. Sci Rep. 2016 Feb 24;6:21780.