KC-2873

293T-LILRA1-Cell-Line

33988

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Background of 293T-LILRA1-Cell-Line

LILRA1 is an activated member of the leukocyte immunoglobulin-like receptor (LIR) family, which is present in the gene cluster in chromosomal region 19q13.4. The encoded protein is primarily expressed in B cells, interacts with major histocompatibility complex class I ligands, and plays a regulatory role in immunosuppressive and immunostimulatory responses. In addition, LIRR is also distributed in certain tumor cells and is associated with malignant tumor progression.Disorders in the expression and function of immunomodulatory molecules (LILRs) may lead to uncontrolled inflammation.

Specifications

Catalog Number	KC-2873
Cell Line Name	293T-LILRA1-Cell-Line
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous LILRA1 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 1:4-1:8 every 2-3 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-LILRA1-cell-line was generated using a lentiviral vector expressing the LILRA1 sequence.

Characterization

Figure 1: Characterization of LILRA1 overexpression in the 293T-LILRA1 stable clone using FACS.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/mL puromycin)in a 37°C water bath. 2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes. 3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol. 4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium. 5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet. 6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask. 7. Incubate the flask at 37°C, 5% CO₂ incubator. 8. Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium. 6. Aliquot 1 mL of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term .

References

1.Hu Y, Lu X, Qiu W, Liu H, Wang Q, Chen Y, Liu W, Feng F, Sun H. The Role of Leukocyte Immunoglobulin-Like Receptors Focusing on the Therapeutic Implications of the Subfamily B2. Curr Drug Targets. 2022;23(15):1430-1452. doi: 10.2174/1389450123666220822201605. PMID: 36017847. 2.Hirayasu K, Arase H. Functional and genetic diversity of leukocyte immunoglobulin-like receptor and implication for disease associations. J Hum Genet. 2015 Nov;60(11):703-8. doi: 10.1038/jhg.2015.64. Epub 2015 Jun 4. PMID: 26040207. 3. Tedla N, An H, Borges L, Vollmer-Conna U, Bryant K, Geczy C, McNeil HP. Expression of activating and inhibitory leukocyte immunoglobulin-like receptors in rheumatoid synovium: correlations to disease activity. Tissue Antigens. 2011 Apr;77(4):305-16. doi: 10.1111/j.1399-0039.2011.01633.x. PMID: 21388353.