KC-1579

293T-LILRB3-Cell-Line

22405

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Background of 293T-LILRB3-Cell-Line

LILRB3, also known as CD85A, is a member of the leukocyte immunoglobulin sample (LIR) family, which is expressed on immune cells and bone marrow, where it binds to MHC class I molecules on antigen-presenting cells and transduces negative signals that inhibit stimulation of the immune response. LILRB3 is thought to control inflammation and cytotoxicity to help focus immune responses and limit autoreactivity. Diseases associated with LILRB3 include Takayasu Arteritis and Anencephaly.

Specifications

Catalog Number	KC-1579
Cell Line Name	293T-LILRB3-Cell-Line
Host Cell Line	Human HEK293T cell line
Description	Stable 293T cell line expressing exogenous human LILRB3 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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10%FBS+1µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 every 3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

293T-LILRB3 cell line was generated using a lentiviral vector expressing the human LILRB3 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 1μg/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 1:3-1:6 every 3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

LILRB3 (ILT5) is a myeloid cell checkpoint that elicits profound immunomodulation.
Allele-specific recognition by LILRB3 and LILRA6 of a cytokeratin 8 - associated ligand on necrotic glandular epithelial cells.
"Entrez Gene: LILRB3 leukocyte immunoglobulin-like receptor, subfamily B (with TM and ITIM domains), member 3".