KC-2337

Ba/F3-LILRB4-Cell-Line

23858

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Background of Ba/F3-LILRB4-Cell-Line

Leukocyte immunoglobulin-like receptor subfamily B member 4 (LILRB4) is also known as CD85 antigen-like family member K (CD85K) or ILT3, which is a member of leukocyte immunoglobulin-like receptor (LIR) family. LILRB4 is detected in monocytes, macrophages, dendritic cells, lung, natural killer cells and B-cells. LILRB4 is receptor for class I MHC antigens. LILRB4 involved in the down-regulation of the immune response and the development of tolerance and inhibits receptor-mediated phosphorylation of cellular proteins and mobilization of intracellular calcium ions. On February 17, 2022, the LILRB4 inhibitor IO-202 received fast track designation from the FDA for the treatment of patients with relapsed or refractory acute myeloid leukemia.

Specifications

Catalog Number	KC-2337
Cell Line Name	Ba/F3-LILRB4-Cell-Line
Host Cell Line	Mouse Ba/F3 cell line
Description	Stable Ba/F3 cell line expressing exogenous human LILRB4 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+1µg/mL Puromycin+8ng/mL mouse IL-3
Selection Marker	Puromycin
Morphology	Mostly single, round (some polymorph) cells in suspension
Subculture	Split saturated culture 1:10 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 20 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Ba/F3 human LILRB4 cell line was generated using a lentiviral vector expressing the human LILRB4 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 1µg/mL Puromycin + 8ng/mL mouse IL-3) 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 1:10 every 3 days; seed out at about 1-3 × 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

Liu J, Wu Q, Shi J, Guo W, Jiang X, Zhou B, Ren C. LILRB4, from the immune system to the disease target. Am J Transl Res. 2020 Jul 15;12(7):3149-3166.
Hammond D, Montalban-Bravo G. Management and Outcomes of Blast Transformed Chronic Myelomonocytic Leukemia. Curr Hematol Malig Rep. 2021 Oct;16(5):405-417.
Stahl M, Goldberg AD. Immune Checkpoint Inhibitors in Acute Myeloid Leukemia: Novel Combinations and Therapeutic Targets. Curr Oncol Rep. 2019 Mar 23;21(4):37.